Parasiticidal compositions comprising indole derivatives, methods and uses thereof

ABSTRACT

The invention relates to oral, topical or injectable compositions for combating liver fluke parasites in mammals, comprising at least one indole derivative active agent. The invention also provides for an improved method for eradicating and controlling liver fluke parasite infections and infestations in a mammal comprising administering the compositions of the invention to the mammal in need thereof.

INCORPORATION BY REFERENCE

This application claims priority to provisional application U.S. Ser.No. 62/014,245, filed on 19 Jun. 2014, and incorporated by referenceherein in its entirety.

FIELD OF THE INVENTION

The present invention provides parasiticidal indole derivatives andoral, topical or injectable veterinary compositions comprising an indolederivative active agent for controlling liver flukes in mammals. The useof these compounds and compositions against liver flukes and methods fortreating parasitic infections and infestations in mammals is presentedherein.

BACKGROUND OF THE INVENTION

Animals such as mammals (including humans) are often susceptible toparasite infections and infestations. These parasites may beectoparasites, such as insects, and endoparasites such as filariae andother worms. Production animals, such as cows, pigs, sheep and goats,can be infected with one or more trematodes. Of particular concern hereis Fasciola hepatica (i.e., liver fluke or F. hepatica).

Liver flukes are a particular problem because they adversely affect thehealth of the animal or human and can inflict significant economic lossin a domestic livestock population. It is estimated that F. hepaticaposes a risk to at least 250 million sheep and 350 million cattleworldwide. Moreover, domestic animals other than sheep and cows mayserve as intermediate hosts. Liver flukes can cause liver condemnation,secondary infections, reduced milk and meat production, abortion andfertility problems.

Several types of control measures for liver flukes have been introducedover the past century. First, halogenated hydrocarbons (e.g., CCl₄;carbon tetrachloride) were introduced for ruminants in the 1920s.Halogenated hydrocarbons had limited success and are no longer usedprimarily because of their adverse effects and variable efficacy.Second, halogenated phenols were administered in the late 1950s (e.g.,hexachlorophene and bithionol sulfoxide) followed by the similarhalogenated salicylanilides (e.g., oxyclozanide, bromoxanide). Fourth,benzimidazole carbamates (e.g., albendazole, luxabendazole) were foundto have a broad anthelmintic spectrum against nematodes and mature F.hepatica. Another benzimidazole—the chlorinated methylthiobenzimidazolederivative triclabendazole—has a high success rate against F. hepatica.Fifth, bisanilino compounds introduced in the 1960s were intolerable dueto toxic side effects. Finally, benzene sulfonamides (e.g., clorsulon)were studied in the 1970s. Extensively modified examples of this classdemonstrate high efficacy on both mature and immature F. hepatica. Ofthese six classes of anthelmintics the benzimidazole class is perhapsthe most widely used for its high efficacy.

Indeed, triclabendazole is the current drug of choice against mature andimmature liver flukes. Not surprisingly, however, reports of parasiteresistance are increasing. For example, Mottier et al., report that apopulation of resistant F. hepatica (Sligo) may use an alteredinflux/efflux mechanism to selectively decrease the amount oftriclabendazole and triclabendazole sulfoxide but not albendazole. SeeMottier et al., J. Parasitol., 92(6), 2006, pp. 1355-1360. McConville etal., report that juvenile triclabendazole-resistant F. hepatica aresomewhat susceptible to compound alpha (i.e.,5-chloro-2-methylthio-6-(1-naphthyloxy)-1H-benzimidazole) via atubulin-independent mechanism. See McConville et al., Parasitol. Res.,(2007) 100:365-377. Further, Keiser et al., report the testing ofartemether and OZ78 in triclabendazole-resistant F. hepatica, althoughat high concentrations. For a short review of triclabendazole resistancesee Brennan et al., Experimental and Molecular Pathology, 82, (2007) pp.104-109.

There is, however, little in the literature disclosing indoles as atreatment for trematodes. Derquantel is in a class known as spiroindolesand is paired with a macrocyclic lactone for greater efficacy againstsheep nematodes (e.g., gastrointestinal and respiratory parasites) butno data is currently available as to the effectiveness of derquantelagainst trematodes.

Other indole compounds have demonstrated an effect on F. hepatica.Indolamines such as serotonin (5-hydroxytryptamine) have a stimulanteffect on the rhythmical activity of the trematode that is antagonizedby other compounds also having an indole structure (e.g., bromolysergicacid diethylamide and yohimbine; Mansour, Brit. J. Pharmacol. (1957),12, 406.)

WO 2007/051619 discloses aryl indole derivatives as pesticides. The '619application speculates that the aryl indole derivatives that itdiscloses would be effective against trematodes such as F. hepatica butoffers evidence only against the dog tick, the cat flea, and certaingastrointestinal nematodes.

WO 2012/125662 discloses indole derivatives useful as ccr2 (i.e.,chemokine receptor type 2) antagonists. The '662 application furtherspeculates that the compounds of its disclosure may be effective againsttrematodes but offers no evidence of such.

Similarly, WO 2012/059232 discloses carbazole and carboline derivatives,and preparation and therapeutic applications thereof. The compoundsreportedly are useful against proliferative diseases and parasiticdiseases. The authors present evidence that some compounds disclosedinhibit tubulin polymerization for possible application againstproliferative diseases but no evidence is presented for activity againsttrematodes.

The resistance to triclabendazole and lack of effective substitutescreates a pressing need in the field for alternatives that exhibit lowside effects and that do not contaminate the animals as a food source.Optimal compositions should further be efficacious, have a quick onsetof activity, have a long duration of activity, and be safe to the animalrecipients and their human owners.

INCORPORATION BY REFERENCE

Any abovementioned applications, and all documents cited therein orduring their prosecution (“application cited documents”) and alldocuments cited or referenced in the application cited documents, andall documents cited or referenced herein (“herein cited documents”), andall documents cited or referenced in herein cited documents, togetherwith any manufacturer's instructions, descriptions, productspecifications, and product sheets for any products mentioned herein orin any document incorporated by reference herein, are herebyincorporated herein by reference, and may be employed in the practice ofthe invention.

Citation or identification of any document in this application is not anadmission that such document is available as prior art to the presentinvention.

SUMMARY OF THE INVENTION

The present invention is directed to compounds and compositions fortreating helminth infestation comprising an anthelmintically effectiveamount of indole compounds of formula (I) described herein and their useto control parasites in mammals to include humans. In accordance withthis invention, it has been discovered that these compounds showunexpected efficacy and speed of onset.

The invention encompasses uses or veterinary uses of the oral, topicalor injectable indole compositions comprising an anthelminticallyeffective amount of indole compounds of formula (I) for the treatment orprophylaxis of parasitic trematode infections and infestations ofanimals (either wild or domesticated), including livestock and companionanimals such as cats, dogs, horses, sheep, goats, pigs and cattle, withthe aim of ridding these hosts of liver flukes encountered by suchanimals. The composition may also be suitable for humans.

The invention also provides methods for treating helminth infestationcomprising administration of an anthelmintically effective amount of thecompound(s) of formula (I) to an animal in need thereof. Surprisingly,it has been found that the inventive compositions and formulationsdescribed herein exhibit superior efficacy against F. hepatica comparedto compositions known in the art.

The invention does not intentionally seek to encompass any previouslyknown product, process of making the product, or method of using theproduct such that the Applicants reserve the right and hereby disclose adisclaimer of any previously known product, process, or method. It isfurther noted that the invention does not intend to encompass within thescope of the invention any product, process, or making of the product ormethod of using the product, which does not meet the written descriptionand enablement requirements of the USPTO (35 U.S.C. §112, firstparagraph) or the EPO (Article 83 of the EPC), such that Applicantsreserve the right and hereby disclose a disclaimer of any previouslydescribed product, process of making the product, or method of using theproduct. The invention and its embodiments are disclosed by thefollowing Detailed Description.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1A shows the effect of various triclabendazole concentrations onliver fluke motility.

FIG. 1B shows the effect of various triclabendazole sulfoxideconcentrations on liver fluke motility.

FIG. 1C shows the effect of various concentrations of compound 332 onliver fluke motility.

FIG. 1D shows the effect of various concentrations of compound 336 onliver fluke motility.

FIG. 1E shows the effect of various concentrations of compound 351 onliver fluke motility.

DETAILED DESCRIPTION

In this disclosure and in the claims, terms such as “comprises,”“comprising,” “containing” and “having” and the like can have themeaning ascribed to them in U.S. Patent law and can mean “includes,”“including,” and the like; “consisting essentially of” or “consistsessentially” likewise has the meaning ascribed in U.S. Patent law andthe term is open-ended, allowing for the presence of more than thatwhich is recited so long as basic or novel characteristics of that whichis recited is not changed by the presence of more than that which isrecited, but excludes prior art embodiments.

It is also noted that in this disclosure and in the claims and/orparagraphs, the compounds of the invention are intended to include allstereoisomers and crystalline forms (which includes hydrated forms,polymorphic forms and amorphous forms with up to 15% by weightcrystalline structure) thereof.

DEFINITIONS

Terms used herein will have their customary meaning in the art unlessspecified otherwise. The organic moieties mentioned in the definitionsof the variables of formula (I) are—like the term halogen—collectiveterms for individual listings of the individual group members. Theprefix C_(n)-C_(m) indicates in each case the possible number of carbonatoms in the group.

The term “animal” is used herein to include all mammals and also includeall vertebrate animals. Animals include, but are not limited to, cats,dogs, cattle, cows, deer, goats, horses, llamas, pigs, sheep and yaks.It also includes an individual animal in all stages of development,including embryonic and fetal stages. In some embodiments, the animalmay be a human animal.

The term “alkyl” refers to saturated straight, branched, cyclic,primary, secondary or tertiary hydrocarbons, including those having 1 to20 atoms. In some embodiments, alkyl groups will include C₁-C₁₂, C₁-C₁₀,C₁-C₈, C₁-C₆ or C₁-C₄ alkyl groups. Examples of C₁-C₁₀ alkyl include,but are not limited to, methyl, ethyl, propyl, 1-methylethyl, butyl,1-methylpropyl, 2-methylpropyl, 1,1-dimethylethyl, pentyl,1-methylbutyl, 2-methylbutyl, 3-methylbutyl, 2,2-dimethylpropyl,1-ethylpropyl, hexyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl,1-methylpentyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl,1,1-dimethylbutyl, 1,2-dimethylbutyl, 1,3-dimethylbutyl,2,2-dimethylbutyl, 2,3-dimethylbutyl, 3,3-dimethylbutyl, 1-ethylbutyl,2-ethylbutyl, 1,1,2-trimethylpropyl, 1,2,2-trimethylpropyl,1-ethyl-1-methylpropyl, 1-ethyl-2-methylpropyl, heptyl, octyl,2-ethylhexyl, nonyl and decyl and their isomers. C₁-C₄-alkyl means forexample methyl, ethyl, propyl, 1-methylethyl, butyl, 1-methylpropyl,2-methylpropyl or 1,1-dimethylethyl.

Cyclic alkyl groups or “cycloalkyl”, which are encompassed by alkylinclude those with 3 to 10 carbon atoms having single or multiplecondensed rings. In some embodiments, cycloalkyl groups include C₄-C₇ orC₃-C₄ cyclic alkyl groups. Non-limiting examples of cycloalkyl groupsinclude adamantyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,cycloheptyl, cyclooctyl and the like.

The alkyl groups described herein can be unsubstituted or substitutedwith one or more moieties selected from the group consisting of alkyl,halo, haloalkyl, hydroxyl, carboxyl, acyl, acyloxy, amino, alkyl- ordialkylamino, amido, arylamino, alkoxy, aryloxy, nitro, cyano, azido,thiol, imino, sulfonic acid, sulfate, sulfonyl, sulfanyl, sulfinyl,sulfamoyl, ester, phosphonyl, phosphinyl, phosphoryl, phosphine,thioester, thioether, acid halide, anhydride, oxime, hydrozine,carbamate, phosphonic acid, phosphate, phosphonate, or any other viablefunctional group that does not inhibit the biological activity of thecompounds of the invention, either unprotected, or protected asnecessary, as known to those skilled in the art, for example, as taughtin Greene, et al., Protective Groups in Organic Synthesis, John Wileyand Sons, Third Edition, 1999, hereby incorporated by reference.

Terms including the term “alkyl” such as “alkylcycloalkyl,”“cycloalkylalkyl,” “alkylamino,” or “dialkylamino” will be understood tocomprise an alkyl group as defined above linked to the other functionalgroup, where the group is linked to the compound through the last grouplisted, as understood by those of skill in the art.

The term “alkenyl” refers to both straight and branched carbon chainswhich have at least one carbon-carbon double bond. In some embodiments,alkenyl groups may include C₂-C₂₀ alkenyl groups. In other embodiments,alkenyl includes C₂-C₁₂, C₂-C₁₀, C₂-C₈, C₂-C₆ or C₂-C₄ alkenyl groups.In one embodiment of alkenyl, the number of double bonds is 1-3, inanother embodiment of alkenyl, the number of double bonds is one or two.Other ranges of carbon-carbon double bonds and carbon numbers are alsocontemplated depending on the location of the alkenyl moiety on themolecule. “C₂-C₁₀-alkenyl” groups may include more than one double bondin the chain. Examples include, but are not limited to, ethenyl,1-propenyl, 2-propenyl, 1-methyl-ethenyl, 1-butenyl, 2-butenyl,3-butenyl, 1-methyl-1-propenyl, 2-methyl-1-propenyl,1-methyl-2-propenyl, 2-methyl-2-propenyl; 1-pentenyl, 2-pentenyl,3-pentenyl, 4-pentenyl, 1-methyl-1-butenyl, 2-methyl-1-butenyl,3-methyl-1-butenyl, 1-methyl-2-butenyl, 2-methyl-2-butenyl,3-methyl-2-butenyl, 1-methyl-3-butenyl, 2-methyl-3-butenyl,3-methyl-3-butenyl, 1,1-dimethyl-2-propenyl, 1,2-dimethyl-1-propenyl,1,2-dimethyl-2-propenyl, 1-ethyl-1-propenyl, 1-ethyl-2-propenyl,1-hexenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl,1-methyl-1-pentenyl, 2-methyl-1-pentenyl, 3-methyl-1-pentenyl,4-methyl-1-pentenyl, 1-methyl-2-pentenyl, 2-methyl-2-pentenyl,3-methyl-2-pentenyl, 4-methyl-2-pentenyl, 1-methyl-3-pentenyl,2-methyl-3-pentenyl, 3-methyl-3-pentenyl, 4-methyl-3-pentenyl,1-methyl-4-pentenyl, 2-methyl-4-pentenyl, 3-methyl-4-pentenyl,4-methyl-4-pentenyl, 1,1-dimethyl-2-butenyl, 1,1-dimethyl-3-butenyl,1,2-dimethyl-1-butenyl, 1,2-dimethyl-2-butenyl, 1,2-dimethyl-3-butenyl,1,3-dimethyl-1-butenyl, 1,3-dimethyl-2-butenyl, 1,3-dimethyl-3-butenyl,2,2-dimethyl-3-butenyl, 2,3-dimethyl-1-butenyl, 2,3-dimethyl-2-butenyl,2,3-dimethyl-3-butenyl, 3,3-dimethyl-1-butenyl, 3,3-dimethyl-2-butenyl,1-ethyl-1-butenyl, 1-ethyl-2-butenyl, 1-ethyl-3-butenyl,2-ethyl-1-butenyl, 2-ethyl-2-butenyl, 2-ethyl-3-butenyl,1,1,2-trimethyl-2-propenyl, 1-ethyl-1-methyl-2-propenyl,1-ethyl-2-methyl-1-propenyl and 1-ethyl-2-methyl-2-propenyl.

“Alkynyl” refers to both straight and branched carbon chains which haveat least one carbon-carbon triple bond. In one embodiment of alkynyl,the number of triple bonds is 1-3; in another embodiment of alkynyl, thenumber of triple bonds is one or two. In some embodiments, alkynylgroups include from C₂-C₂₀ alkynyl groups. In other embodiments, alkynylgroups may include C₂-C₁₂, C₂-C₁₀, C₂-C₈, C₂-C₆ or C₂-C₄ alkynyl groups.Other ranges of carbon-carbon triple bonds and carbon numbers are alsocontemplated depending on the location of the alkynyl moiety on themolecule. For example, the term “C₂-C₁₀-alkynyl” as used herein refersto a straight-chain or branched unsaturated hydrocarbon group having 2to 10 carbon atoms and containing at least one triple bond, such asethynyl, prop-1-yn-1-yl, prop-2-yn-1-yl, n-but-1-yn-1-yl,n-but-1-yn-3-yl, n-but-1-yn-4-yl, n-but-2-yn-1-yl, n-pent-1-yn-1-yl,n-pent-1-yn-3-yl, n-pent-1-yn-4-yl, n-pent-1-yn-5-yl, n-pent-2-yn-1-yl,n-pent-2-yn-4-yl, n-pent-2-yn-5-yl, 3-methylbut-1-yn-3-yl,3-methylbut-1-yn-4-yl, n-hex-1-yn-1-yl, n-hex-1-yn-3-yl,n-hex-1-yn-4-yl, n-hex-1-yn-5-yl, n-hex-1-yn-6-yl, n-hex-2-yn-1-yl,n-hex-2-yn-4-yl, n-hex-2-yn-5-yl, n-hex-2-yn-6-yl, n-hex-3-yn-1-yl,n-hex-3-yn-2-yl, 3-methylpent-1-yn-1-yl, 3-methylpent-1-yn-3-yl,3-methylpent-1-yn-4-yl, 3-methylpent-1-yn-5-yl, 4-methylpent-1-yn-1-yl,4-methylpent-2-yn-4-yl or 4-methylpent-2-yn-5-yl and the like.

The term “haloalkyl” refers to an alkyl group, as defined herein, whichis substituted by one or more halogen atoms. For example C₁-C₄-haloalkylincludes, but is not limited to, chloromethyl, bromomethyl,dichloromethyl, trichloromethyl, fluoromethyl, difluoromethyl,trifluoromethyl, chlorofluoromethyl, dichlorofluoromethyl,chlorodifluoromethyl, 1-chloroethyl, 1-bromoethyl, 1-fluoroethyl,2-fluoroethyl, 2,2-difluoroethyl, 2,2,2-trifluoroethyl,2-chloro-2-fluoroethyl, 2-chloro-2,2-difluoroethyl,2,2-dichloro-2-fluoroethyl, 2,2,2-trichloroethyl, pentafluoroethyl andthe like.

The term “haloalkenyl” refers to an alkenyl group, as defined herein,which is substituted by one or more halogen atoms.

The term “haloalkynyl” refers to an alkynyl group, as defined herein,which is substituted by one or more halogen atoms.

“Alkoxy” refers to alkyl-O—, wherein alkyl is as defined above.Similarly, the terms “alkenyloxy,” “alkynyloxy,” “haloalkoxy,”“haloalkenyloxy,” “haloalkynyloxy,” “cycloalkoxy,” “cycloalkenyloxy,”“halocycloalkoxy,” and “halocycloalkenyloxy” refer to the groupsalkenyl-O—, alkynyl-O—, haloalkyl-O—, haloalkenyl-O—, haloalkynyl-O—,cycloalkyl-O—, cycloalkenyl-O—, halocycloalkyl-O—, andhalocycloalkenyl-O—, respectively, wherein alkenyl, alkynyl, haloalkyl,haloalkenyl, haloalkynyl, cycloalkyl, cycloalkenyl, halocycloalkyl, andhalocycloalkenyl are as defined above. Examples of C₁-C₆-alkoxy include,but are not limited to, methoxy, ethoxy, C₂H₅—CH₂O—, (CH₃)₂CHO—,n-butoxy, C₂H₅—CH(CH₃)O—, (CH₃)₂CH—CH₂O—, (CH₃)₃CO—, n-pentoxy,1-methylbutoxy, 2-methylbutoxy, 3-methylbutoxy, 1,1-dimethylpropoxy,1,2-dimethylpropoxy, 2,2-dimethyl-propoxy, 1-ethylpropoxy, n-hexoxy,1-methylpentoxy, 2-methylpentoxy, 3-methylpentoxy, 4-methylpentoxy,1,1-dimethylbutoxy, 1,2-dimethylbutoxy, 1,3-dimethylbutoxy,2,2-dimethylbutoxy, 2,3-dimethylbutoxy, 3,3-dimethylbutoxy,1-ethylbutoxy, 2-ethylbutoxy, 1,1,2-trimethylpropoxy,1,2,2-trimethylpropoxy, 1-ethyl-1-methylpropoxy, 1-ethyl-2-methylpropoxyand the like.

The term “alkylthio” refers to alkyl-S—, wherein alkyl is as definedabove. Similarly, the terms “haloalkylthio,” “cycloalkylthio,” and thelike, refer to haloalkyl-S— and cycloalkyl-S— where haloalkyl andcycloalkyl are as defined above.

The term “alkylsulfinyl” refers to alkyl-S(O)—, wherein alkyl is asdefined above. Similarly, the term “haloalkylsulfinyl” refers tohaloalkyl-S(O)— where haloalkyl is as defined above.

The term “alkylsulfonyl” refers to alkyl-S(O)₂—, wherein alkyl is asdefined above. Similarly, the term “haloalkylsulfonyl” refers tohaloalkyl-S(O)₂— where haloalkyl is as defined above.

The term alkylamino and dialkylamino refer to alkyl-NH— and (alkyl)₂N—where alkyl is as defined above. Similarly, the terms “haloalkylamino”refers to haloalkyl-NH— where haloalkyl is as defined above.

The terms “alkylcarbonyl,” “alkoxycarbonyl,” “alkylaminocarbonyl,” and“dialkylaminocarbonyl refer to alkyl-C(O)—, alkoxy-C(O)—,alkylamino-C(O)— and dialkylamino-C(O)— where alkyl, alkoxy, alkylaminoand dialkylamino are as defined above. Similarly, the terms“haloalkylcarbonyl,” “haloalkoxycarbonyl,” “haloalkylaminocarbonyl,” and“dihaloalkylaminocarbonyl” refer to the groups haloalkyl-C(O)—,haloalkoxy-C(O)—, haloalkylamino-C(O)— and dihaloalkylamino-C(O)— wherehaloalkyl, haloalkoxy, haloalkylamino and dihaloalkylamino are asdefined above.

“Aryl” refers to a monovalent aromatic carbocyclic group of from 6 to 14carbon atoms having a single ring or multiple condensed rings. In someembodiments, aryl groups include C₆-C₁₀ aryl groups. Aryl groupsinclude, but are not limited to, phenyl, biphenyl, naphthyl,tetrahydronaphthyl, phenylcyclopropyl and indanyl. Aryl groups may beunsubstituted or substituted by one or more moieties selected fromhalogen, cyano, nitro, hydroxy, mercapto, amino, alkyl, alkenyl,alkynyl, cycloalkyl, cycloalkenyl, haloalkyl, haloalkenyl, haloalkynyl,halocycloalkyl, halocycloalkenyl, alkoxy, alkenyloxy, alkynyloxy,haloalkoxy, haloalkenyloxy, haloalkynyloxy, cycloalkoxy,cycloalkenyloxy, halocycloalkoxy, halocycloalkenyloxy, alkylthio,haloalkylthio, cycloalkylthio, halocycloalkylthio, alkylsulfinyl,alkenylsulfinyl, alkynyl-sulfinyl, haloalkylsulfinyl,haloalkenylsulfinyl, haloalkynylsulfinyl, alkylsulfonyl,alkenylsulfonyl, alkynylsulfonyl, haloalkyl-sulfonyl,haloalkenylsulfonyl, haloalkynylsulfonyl, alkylamino, alkenylamino,alkynylamino, di(alkyl)amino, di(alkenyl)-amino, di(alkynyl)amino, ortrialkylsilyl. Aryl groups may be linked to other moieties via an etherlinkage.

The term “aralkyl” refers to an aryl group that is bonded to the parentcompound through a diradical alkylene bridge, (—CH₂—)—, where n is 1-12and where “aryl” is as defined above.

“Heteroaryl” refers to a monovalent aromatic group of from 1 to 15carbon atoms, such as from 1 to 10 carbon atoms, having one or moreoxygen, nitrogen, and sulfur heteroatoms within the ring, for example 1to 4 heteroatoms, or 1 to 3 heteroatoms. The nitrogen and sulfurheteroatoms may optionally be oxidized. Such heteroaryl groups can havea single ring (e.g., pyridyl or furyl) or multiple condensed ringsprovided that the point of attachment is through a heteroaryl ring atom.Heteroaryls may include pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl,triazinyl, pyrrolyl, indolyl, quinolinyl, isoquinolinyl, quinazolinyl,quinoxalinyl, furanyl, thiophenyl, furyl, pyrrolyl, imidazolyl,oxazolyl, isoxazolyl, isothiazolyl, pyrazolyl benzofuranyl, andbenzothiophenyl. Heteroaryl rings may be unsubstituted or substituted byone or more moieties as described for aryl above. Heteroaryl groups maybe linked to other moieties via an ether linkage.

“Heterocyclyl,” “heterocyclic” or “heterocyclo” refer to fully saturatedor unsaturated, cyclic groups, for example, 3 to 7 membered monocyclicor 4 to 7 membered monocyclic; 7 to 11 membered bicyclic, or 10 to 15membered tricyclic ring systems, which have one or more oxygen, sulfuror nitrogen heteroatoms in ring, such as 1 to 4 or 1 to 3 heteroatoms.The nitrogen and sulfur heteroatoms may optionally be oxidized and thenitrogen heteroatoms may optionally be quaternized. The heterocyclicgroup may be attached at any heteroatom or carbon atom of the ring orring system and may be unsubstituted or substituted by one or moremoieties as described for aryl groups above.

Exemplary monocyclic heterocyclic groups include, but are not limitedto, pyrrolidinyl, pyrrolyl, pyrazolyl, oxetanyl, pyrazolinyl,imidazolyl, imidazolinyl, imidazolidinyl, oxazolyl, oxazolidinyl,isoxazolinyl, isoxazolyl, thiazolyl, thiadiazolyl, thiazolidinyl,isothiazolyl, isothiazolidinyl, furyl, tetrahydrofuryl, thienyl,oxadiazolyl, piperidinyl, piperazinyl, 2-oxopiperazinyl,2-oxopiperidinyl, 2-oxopyrrolidinyl, 2-oxoazepinyl, azepinyl,4-piperidonyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl,tetrahydropyranyl, morpholinyl, thiamorpholinyl, thiamorpholinylsulfoxide, thiamorpholinyl sulfone, 1,3-dioxolane andtetrahydro-1,1-dioxothienyl, triazolyl, triazinyl, and the like.

Exemplary bicyclic heterocyclic groups include, but are not limited to,indolyl, benzothiazolyl, benzoxazolyl, benzodioxolyl, benzothienyl,quinuclidinyl, quinolinyl, tetra-hydroisoquinolinyl, isoquinolinyl,benzimidazolyl, benzopyranyl, indolizinyl, benzofuryl, chromonyl,coumarinyl, benzopyranyl, cinnolinyl, quinoxalinyl, indazolyl,pyrrolopyridyl, furopyridinyl (such as furo[2,3-c]pyridinyl,furo[3,2-b]pyridinyl]or furo[2,3-b]pyridinyl), dihydroisoindolyl,dihydroquinazolinyl (such as 3,4-dihydro-4-oxo-quinazolinyl),tetrahydroquinolinyl and the like.

Exemplary tricyclic heterocyclic groups include carbazolyl, benzidolyl,phenanthrolinyl, acridinyl, phenanthridinyl, xanthenyl, and the like.

Halogen means the atoms fluorine, chlorine, bromine and iodine. Thedesignation of “halo” (e.g., as illustrated in the term haloalkyl)refers to all degrees of substitutions from a single substitution to aperhalo substitution (e.g., as illustrated with methyl as chloromethyl(—CH₂Cl), dichloromethyl (—CHCl₂), trichloromethyl (—CCl₃)).

Stereoisomers and Polymorphic Forms

It will be appreciated by those of skill in the art that certaincompounds within the compositions of the invention may exist and beisolated as optically active and racemic forms. Compounds having one ormore chiral centers, including at a sulfur atom, may be present assingle enantiomers or diastereomers or as mixtures of enantiomers and/ordiastereomers. For example, it is well known in the art that sulfoxidecompounds may be optically active and may exist as single enantiomers orracemic mixtures. In addition, compounds within the compositions of theinvention may include one or more chiral centers, which results in atheoretical number of optically active isomers. Where compounds withinthe compositions of the invention include “n” chiral centers, thecompounds may comprise up to 2^(n) optical isomers. The presentinvention encompasses the specific enantiomers or diastereomers of eachcompound as well as mixtures of different enantiomers and/ordiastereomers of the compounds of the invention that possess the usefulproperties described herein. The optically active forms can be preparedby, for example, resolution of the racemic forms by selectivecrystallization techniques, by synthesis from optically activeprecursors, by chiral synthesis, by chromatographic separation using achiral stationary phase or by enzymatic resolution.

The compounds within the compositions of the present invention may alsobe present in different solid forms such as different crystalline formsor in the form of an amorphous solid. The present invention encompassesdifferent crystalline forms as well as amorphous forms of the inventivecompounds.

In addition, the compounds within the compositions of the invention mayexist as hydrates or solvates, in which a certain stoichiometric amountof water or a solvent is associated with the molecule in the crystallineform. The compositions of the invention may include hydrates andsolvates of the active agents.

Salts

Also contemplated within the scope of the invention are acid or basesalts, where applicable, of the compounds of the invention provided forherein.

The term “acid” contemplates all pharmaceutically acceptable inorganicor organic acids. Inorganic acids include mineral acids such ashydrohalic acids such as hydrobromic acid and hydrochloric acid,sulfuric acid, phosphoric acids and nitric acid. Organic acids includeall pharmaceutically acceptable aliphatic, alicyclic and aromaticcarboxylic acids, dicarboxylic acids, tricarboxylic acids and fattyacids. In one embodiment of the acids, the acids are straight chain orbranched, saturated or unsaturated C₁-C₂₀ aliphatic carboxylic acids,which are optionally substituted by halogen or by hydroxyl groups, orC₆-C₁₂ aromatic carboxylic acids. Examples of such acids are carbonicacid, formic acid, acetic acid, propionic acid, isopropionic acid,valeric acid, α-hydroxy acids such as glycolic acid and lactic acid,chloroacetic acid, benzoic acid, methane sulfonic acid, and salicylicacid. Examples of dicarboxylic acids include oxalic acid, malic acid,succinic acid, tartaric acid, fumaric acid, and maleic acid. An exampleof a tricarboxylic acid is citric acid. Fatty acids include allpharmaceutically acceptable saturated or unsaturated aliphatic oraromatic carboxylic acids having 4 to 24 carbon atoms. Examples includebutyric acid, isobutyric acid, sec-butyric acid, lauric acid, palmiticacid, stearic acid, oleic acid, linoleic acid, linolenic acid, andphenylstearic acid. Other acids include gluconic acid, glycoheptonicacid and lactobionic acid.

The term “base” contemplates all pharmaceutically acceptable inorganicor organic bases, including hydroxides, carbonates or bicarbonates ofalkali metal or alkaline earth metals. Salts formed with such basesinclude, for example, the alkali metal and alkaline earth metal salts,including, but not limited to, as the lithium, sodium, potassium,magnesium or calcium salts. Salts formed with organic bases include thecommon hydrocarbon and heterocyclic amine salts, which include, forexample, ammonium salts (NH₄ ⁺), alkyl- and dialkylammonium salts, andsalts of cyclic amines such as the morpholine and piperidine salts.

The term “derivative” contemplates a compound obtained from or closelyrelated to another substance or compound. A derivative contemplates achemical compound that may be produced from another chemical compound ofsimilar structure in one or more steps.

In one embodiment, the invention provides novel oral, topical orinjectable veterinary compounds according to formula (I) below.

wherein:

-   -   R₁ is (C₁-C₆)-alkyl, (C₂-C₅)-alkenyl, or (C₂-C₅)-alkynyl, each        independently unsubstituted or substituted with one or more        halogens; R₂ is H, halogen, (C₁-C₆)-alkyl or (C₁-C₆) haloalkyl;        R₃ is halogen, (C₁-C₆)-alkyl, (C₂-C₅)-alkenyl, or        (C₂-C₅)-alkynyl, (C₁-C₆)-alkoxy, thienyl, furanyl, biphenyl,        naphthyl, tetralin ether, aryl, aryl ether, sulfonylaryl,        sulfoxyaryl, thioaryl, (C₃-C₆)-cycloalkyl, or pyridinyl, wherein        the last seven substituents may be unsubstituted or substituted        with one or more of halogen, (C₁-C₃)-haloalkyl,        (C₁-C₃)-haloalkoxy or carboxyl; R₄ is H, halogen, (C₁-C₆)-alkyl        or (C₁-C₆)-haloalkyl; R₅ is H, halogen, (C₁-C₆)-alkyl or        (C₁-C₆)-haloalkyl; and R₆ is H, halogen, (C₁-C₆)-alkyl,        (C₂-C₅)-alkenyl, or (C₂-C₅)-alkynyl, wherein each of        (C₁-C₆)-alkyl, (C₂-C₅)-alkenyl, or (C₂-C₅)-alkynyl is        independently unsubstituted or substituted with one or more        halogens.

In another embodiment, the compound of formula (I) defines R₁ as(C₁-C₄)-alkyl substituted with fluorine. In yet another embodiment, thecompound of formula (I) defines R₂ as chlorine.

In another embodiment, the compound of formula (I) defines R₃ as a mono-or di-halo substituted phenyl ether. In another embodiment, the compoundof formula (I) defines R₃ as a di- or tri-halo substituted phenyl. Inyet another embodiment, the compound of formula (I) defines R₃ aschlorine, bromine or fluorine.

In another embodiment, the compound of formula (I) defines R₄ aschlorine. In another embodiment, the compound of formula (I) defines R₅as hydrogen.

In another embodiment, the compound of formula (I) is5-[2-chloro-4-(trifluoromethyl)phenyl]-2-(trifluoromethyl)-1H-indole. Inanother embodiment, the compound of formula (I) is6-[2-chloro-4-(trifluoromethyl)phenyl]-2-(trifluoromethyl)-1H-indole. Inanother embodiment, the compound of formula (I) is6-chloro-5-[2-chloro-4-(trifluoromethyl)phenyl]-2-(trifluoromethyl)-1H-indole.In another embodiment, the compound of formula (I) is6-chloro-2-(trifluoromethyl)-5-[4-(trifluoromethyl)phenyl]-1H-indole. Inanother embodiment, the compound of formula (I) is6-chloro-5-(2-chlorophenyl)-2-(trifluoromethyl)-1H-indole.

In another aspect, the invention is a composition for treating helminthinfestation comprising an anthelmintically effective amount of thecompounds of formula (I) defined above and a pharmaceutically acceptablecarrier.

In another embodiment, the composition including formula (I) is combinedwith a macrocyclic lactone. In one embodiment, the macrocyclic lactoneis an avermectin. In another embodiment, the macrocyclic lactone isivermectin.

In yet another embodiment, the composition including formula (I) iscombined with verapamil. Verapamil and macrocyclic lactones such asavermectins may provide a synergistic effect in combination withcompounds of formula (I) to kill triclabendazole-resistant liver flukes.

The synergistic effect is thought to occur due to the inhibition of theP-glycoprotein drug transporter by verapamil or the avermectin.

In another aspect, the invention is a method for treating helminthinfestation comprising administration of an anthelmintically effectiveamount of the compounds defined above to an animal in need thereof. Thehelminths are, for example, trematodes, and may be specifically F.hepatica.

Experimental Procedure and Results

Representative compounds of formula (I) were tested in vitro against F.hepatica as follows. Adult F. hepatica (fluke) were collected frominfected bovine livers obtained from a local abattoir (Basel,Switzerland). The worms were quickly washed with 0.9% (w/v) NaCl andplaced in 6 or 12-well plates (Costar). Culture medium in each wellcontained RPMI 1640 (Gibco) at 37° C., which was supplemented withantibiotics (50 μg/ml streptomycin and 50 IU/ml penicillin; Gibco) and80 μg/ml of a haemin solution. The haemin solution was prepared asfollows: 5 mg haemin was dissolved in 1 ml of 0.1M aqueous solution ofNaOH, and 3.95 ml of PBS (pH=7.4) and 0.05 ml of 1M HCl were added toadjust the pH to 7.1-7.4 (Keiser and Morson, 2008). Cultures were keptat 37° C. in an atmosphere of 5% CO₂. To monitor the temporal effects oftest compounds in vitro, 3 flukes were incubated for 72 h in thepresence of 50 or 100 μg/ml of each test compound. At 24, 48, and 72 h,worms were examined using a dissecting microscope. For the adult worms,a viability scale ranging from 3 (normal movements) to 0 (death; nomovement observed for two min using a microscope) was used. Testcompounds that showed activity at a concentration of 50 μg/ml werefurther evaluated at lower concentrations (20 μg/ml, 10 μg/ml, 5 μg/ml,2.5 μg/ml, 1.25 μg/ml and 0.625 μg/ml).

Table 1 lists representative compounds of formula (I) of the presentinvention and their effective in vitro concentrations against F.hepatica. All compounds were tested in vitro on the adult stage of F.hepatica as described above. Each data point refers to at least twoindependent experiments having at least three worms per concentration.

TABLE 1 Effective in vitro Com- concen- pound tration; No. Structureμg/mL 332

<10 334

<10 336

<10 337

<10 338

<10 339

<10 340

<10 341

>10 342

<10 343

<10 344

<10 345

<10 346

<10 347

≦10 348

<10 349

≦10 350

≦10 351

<10 352

>10 354

>2.5 355

≧2.5 356

<10 357

<10 358

>2.5 359

>2.5 360

>2.5 360-0A

>2.5 361

<10 361-0A

<10 362

>2.5 362-0A

>2.5

Further testing employed a fluke motility assay. Compounds 332, 336 and351 were screened for activity against liver fluke newly-excystedjuveniles (NEJs) in a digital image-based motility assay, based onmeasurement of the rate of change in worm length (μm/min) duringperistaltic motility. Newly excised worms were immediately placed into asolution of drug in RPMI (final concentrations 10 μM-1 nM), alongsideuntreated and vehicle controls, and triclabendazole/triclabendazolesulfoxide (i.e., TCBZ/TCBZ-SO) at matching concentrations, for a periodof 18 h. Each treatment group consisted of 15-20 NEJs. Compounds 332(FIG. 1C), 336 (FIG. 1D) and 351 (FIG. 1E) showed more potent inhibitionof motility than either TCBZ (FIG. 1A) or TCBZ-SO (FIG. 1B; botheffective only at 10 μM). Compounds 332 and 351 completely abolishedmotility at 1 μM, while compound 336 significantly impacted motility at100 nM. See FIG. 1. Each data point represents mean length change(μm/min) in an individual worm during a five minute recording period.Horizontal lines represent data set means. ****, p<0.0001; ***, p<0.001;**, p<0.01; *, p<0.05.

Additional veterinary/pharmaceutical active ingredients may be used withthe compositions of the invention for oral, topical or injectable use.In some embodiments, the additional active agents may include, but arenot limited to, acaricides, anthelmintics, anti-parasitics andinsecticides. Anti-parasitic agents can include both ectoparasiticidaland endoparasiticidal agents.

Veterinary pharmaceutical agents that may be included in thecompositions of the invention are well-known in the art (see e.g.,Plumb' Veterinary Drug Handbook, 5^(th) Edition, ed. Donald C. Plumb,Blackwell Publishing, (2005) or The Merck Veterinary Manual, 9^(th)Edition, (January 2005)) and include but are not limited to acarbose,acepromazine maleate, acetaminophen, acetazolamide, acetazolamidesodium, acetic acid, acetohydroxamic acid, acetylcysteine, acitretin,acyclovir, albendazole, albuterol sulfate, alfentanil, allopurinol,alprazolam, altrenogest, amantadine, amikacin sulfate, aminocaproicacid, aminopentamide hydrogen sulfate, aminophylline/theophylline,amiodarone, amitriptyline, amlodipine besylate, ammonium chloride,ammonium molybdenate, amoxicillin, clavulanate potassium, amphotericin Bdesoxycholate, amphotericin B lipid-based, ampicillin, amprolium,antacids (oral), antivenin, apomorphione, apramycin sulfate, ascorbicacid, asparaginase, aspiring, atenolol, atipamezole, atracuriumbesylate, atropine sulfate, aurnofin, aurothioglucose, azaperone,azathioprine, azithromycin, baclofen, barbituates, benazepril,betamethasone, bethanechol chloride, bisacodyl, bismuth subsalicylate,bleomycin sulfate, boldenone undecylenate, bromides, bromocriptinemesylate, budenoside, buprenorphine, buspirone, busulfan, butorphanoltartrate, cabergoline, calcitonin salmon, calcitrol, calcium salts,captopril, carbenicillin indanyl sodium, carbimazole, carboplatin,carnitine, carprofen, carvedilol, cefadroxil, cefazolin sodium,cefixime, chlorsulon, cefoperazone sodium, cefotaxime sodium, cefotetandisodium, cefoxitin sodium, cefpodoxime proxetil, ceftazidime, ceftiofursodium, ceftiofur, ceftiaxone sodium, cephalexin, cephalosporins,cephapirin, charcoal (activated), chlorambucil, chloramphenicol,chlordiazepoxide, chlordiazepoxide+/−clidinium bromide, chlorothiazide,chlorpheniramine maleate, chlorpromazine, chlorpropamide,chlortetracycline, chorionic gonadotropin (HCG), chromium, cimetidine,ciprofloxacin, cisapride, cisplatin, citrate salts, clarithromycin,clemastine fumarate, clenbuterol, clindamycin, clofazimine,clomipramine, claonazepam, clonidine, cloprostenol sodium, clorazepatedipotassium, clorsulon, cloxacillin, codeine phosphate, colchicine,corticotropin (ACTH), cosyntropin, cyclophosphamide, cyclosporine,cyproheptadine, cytarabine, dacarbazine, dactinomycin/actinomycin D,dalteparin sodium, danazol, dantrolene sodium, dapsone, decoquinate,deferoxamine mesylate, deracoxib, deslorelin acetate, desmopressinacetate, desoxycorticosterone pivalate, detomidine, dexamethasone,dexpanthenol, dexraazoxane, dextran, diazepam, diazoxide (oral),dichlorphenamide, diclofenac sodium, dicloxacillin, diethylcarbamazinecitrate, diethylstilbestrol (DES), difloxacin, digoxin,dihydrotachysterol (DHT), diltiazem, dimenhydrinate, dimercaprol/BAL,dimethyl sulfoxide, dinoprost tromethamine, diphenylhydramine,disopyramide phosphate, dobutamine, docusate/DSS, dolasetron mesylate,domperidone, dopamine, doramectin, doxapram, doxepin, doxorubicin,doxycycline, edetate calcium disodium.calcium EDTA, edrophoniumchloride, enalapril/enalaprilat, enoxaparin sodium, enrofloxacin,ephedrine sulfate, epinephrine, epoetin/erythropoietin, eprinomectin,epsiprantel, erythromycin, esmolol, estradiol cypionate, ethacrynicacid/ethacrynate sodium, ethanol (alcohol), etidronate sodium, etodolac,etomidate, euthanasia agents w/pentobarbital, famotidine, fatty acids(essential/omega), felbamate, fentanyl, ferrous sulfate, filgrastim,finasteride, fipronil, florfenicol, fluconazole, flucytosine,fludrocortisone acetate, flumazenil, flumethasone, flunixin meglumine,fluorouracil (5-FU), fluoxetine, fluticasone propionate, fluvoxaminemaleate, fomepizole (4-MP), furazolidone, furosemide, gabapentin,gemcitabine, gentamicin sulfate, glimepiride, glipizide, glucagon,glucocorticoid agents, glucosamine/chondroitin sulfate, glutamine,glyburide, glycerine (oral), glycopyrrolate, gonadorelin, grisseofulvin,guaifenesin, halothane, hemoglobin glutamer-200 (Oxyglobin®), heparin,hetastarch, hyaluronate sodium, hydrazaline, hydrochlorothiazide,hydrocodone bitartrate, hydrocortisone, hydromorphone, hydroxyurea,hydroxyzine, ifosfamide, imidacloprid, imidocarb dipropinate,impenem-cilastatin sodium, imipramine, inamrinone lactate, insulin,interferon alfa-2a (human recombinant), iodide (sodium/potassium),ipecac (syrup), ipodate sodium, iron dextran, isoflurane, isoproterenol,isotretinoin, isoxsuprine, itraconazole, ivermectin, kaolin/pectin,ketamine, ketoconazole, ketoprofen, ketorolac tromethamine, lactulose,leuprolide, levamisole, levetiracetam, levothyroxine sodium, lidocaine,lincomycin, liothyronine sodium, lisinopril, lomustine (CCNU),lufenuron, lysine, magnesium, mannitol, marbofloxacin, mechlorethamine,meclizine, meclofenamic acid, medetomidine, medium chain triglycerides,medroxyprogesterone acetate, megestrol acetate, melarsomine, melatonin,meloxican, melphalan, meperidine, mercaptopurine, meropenem, metformin,methadone, methazolamide, methenamine mandelate/hippurate, methimazole,methionine, methocarbamol, methohexital sodium, methotrexate,methoxyflurane, methylene blue, methylphenidate, methylprednisolone,metoclopramide, metoprolol, metronidaxole, mexiletine, mibolerlone,midazolam milbemycin oxime, mineral oil, minocycline, misoprostol,mitotane, mitoxantrone, morphine sulfate, moxidectin, naloxone,mandrolone decanoate, naproxen, narcotic (opiate) agonist analgesics,neomycin sulfate, neostigmine, niacinamide, nitazoxanide, nitenpyram,nitrofurantoin, nitroglycerin, nitroprusside sodium, nizatidine,novobiocin sodium, nystatin, octreotide acetate, olsalazine sodium,omeprozole, ondansetron, opiate antidiarrheals, orbifloxacin, oxacillinsodium, oxazepam, oxibutynin chloride, oxymorphone, oxytretracycline,oxytocin, pamidronate disodium, pancreplipase, pancuronium bromide,paromomycin sulfate, parozetine, pencillamine, general informationpenicillins, penicillin G, penicillin V potassium, pentazocine,pentobarbital sodium, pentosan polysulfate sodium, pentoxifylline,pergolide mesylate, phenobarbital, phenoxybenzamine, pheylbutazone,phenylephrine, phenypropanolamine, phenytoin sodium, pheromones,parenteral phosphate, phytonadione/vitamin K-1, pimobendan, piperazine,pirlimycin, piroxicam, polysulfated glycosaminoglycan, ponazuril,potassium chloride, pralidoxime chloride, prazosin,prednisolone/prednisone, primidone, procainamide, procarbazine,prochlorperazine, propantheline bromide, propionibacterium acnesinjection, propofol, propranolol, protamine sulfate, pseudoephedrine,psyllium hydrophilic mucilloid, pyridostigmine bromide, pyrilaminemaleate, pyrimethamine, quinacrine, quinidine, ranitidine, rifampin,s-adenosyl-methionine (SAMe), saline/hyperosmotic laxative, selamectin,selegiline/l-deprenyl, sertraline, sevelamer, sevoflurane,silymarin/milk thistle, sodium bicarbonate, sodium polystyrenesulfonate, sodium stibogluconate, sodium sulfate, sodum thiosulfate,somatotropin, sotalol, spectinomycin, spironolactone, stanozolol,streptokinase, streptozocin, succimer, succinylcholine chloride,sucralfate, sufentanil citrate, sulfachlorpyridazine sodium,sulfadiazine/trimethroprim, sulfamethoxazole/trimethoprim,sulfadimentoxine, sulfadimethoxine/ormetoprim, sulfasalazine, taurine,tepoxaline, terbinafline, terbutaline sulfate, testosterone,tetracycline, thiacetarsamide sodium, thiamine, thioguanine, thiopentalsodium, thiotepa, thyrotropin, tiamulin, ticarcilin disodium,tiletamine/zolazepam, tilmocsin, tiopronin, tobramycin sulfate,tocainide, tolazoline, telfenamic acid, topiramate, tramadol,trimcinolone acetonide, trientine, trilostane, trimepraxine tartratew/prednisolone, tripelennamine, tylosin, urdosiol, valproic acid,vanadium, vancomycin, vasopressin, vecuronium bromide, verapamil,vinblastine sulfate, vincristine sulfate, vitamin E/selenium, warfarinsodium, xylazine, yohimbine, zafirlukast, zidovudine (AZT), zincacetate/zinc sulfate, zonisamide and mixtures thereof.

In another embodiment of the invention, one or more macrocyclic lactonesor lactams, which act as an acaricide, anthelmintic agent and/orinsecticide, can be added to the compositions of the invention.

The macrocyclic lactones include, but are not limited to, avermectins,such as abamectin, dimadectin, doramectin, emamectin, eprinomectin,ivermectin, latidectin, lepimectin, selamectin, ML-1,694,554 andmilbemycins, such as milbemectin, milbemycin D, moxidectin andnemadectin. Also included are the 5-oxo and 5-oxime derivatives of saidavermectins and milbemycins.

The macrocyclic lactone compounds are known in the art and can easily beobtained commercially or through synthesis techniques known in the art.Reference is made to the widely available technical and commercialliterature. For avermectins, ivermectin and abamectin, reference may bemade, for example, to the work “Ivermectin and Abamectin”, 1989, by M.H. Fischer and H. Mrozik, William C. Campbell, published by SpringerVerlag., or Albers-Schönberg et al. (1981), “Avermectins StructureDetermination”, J. Am. Chem. Soc., 103, 4216-4221. For doramectin,“Veterinary Parasitology”, vol. 49, No. 1, July 1993, 5-15 may beconsulted. For milbemycins, reference may be made, inter alia, to DaviesH. G. et al., 1986, “Avermectins and Milbemycins”, Nat. Prod. Rep., 3,87-121, Mrozik H. et al., 1983, Synthesis of Milbemycins fromAvermectins, Tetrahedron Lett., 24, 5333-5336, U.S. Pat. No. 4,134,973and EP 0 677 054.

Macrocyclic lactones are either natural products or are semi-syntheticderivatives thereof. The structure of the avermectins and milbemycinsare closely related, e.g., by sharing a complex 16-membered macrocycliclactone ring. The natural product avermectins are disclosed in U.S. Pat.No. 4,310,519 and the 22,23-dihydro avermectin compounds are disclosedin U.S. Pat. No. 4,199,569. Mention is also made of U.S. Pat. Nos.4,468,390, 5,824,653, EP 0 007 812 A1, U.K. Patent Specification 1 390336, EP 0 002 916, and New Zealand Patent No. 237 086, inter alia.Naturally occurring milbemycins are described in U.S. Pat. No. 3,950,360as well as in the various references cited in “The Merck Index” 12^(th)ed., S. Budavari, Ed., Merck & Co., Inc. Whitehouse Station, N.J.(1996). Latidectin is described in the “International NonproprietaryNames for Pharmaceutical Substances (INN)”, WHO Drug Information, vol.17, no. 4, pp. 263-286, (2003). Semisynthetic derivatives of theseclasses of compounds are well known in the art and are described, forexample, in U.S. Pat. Nos. 5,077,308, 4,859,657, 4,963,582, 4,855,317,4,871,719, 4,874,749, 4,427,663, 4,310,519, 4,199,569, 5,055,596,4,973,711, 4,978,677, 4,920,148 and EP 0 667 054.

In another embodiment, the invention comprises a composition comprisingan indole compound in combination with verapamil. Verapamil is believedto be an inhibitor of P-glycoprotein, which is a membrane protein thathas been shown to efflux triclabendazole from triclabendazole-resistantF. hepatica. Inhibiting the efflux mechanism could allow thebenzimidazole derivative to accumulate to toxic levels in the parasite.

In another embodiment, the invention comprises a composition comprisingan indole compound in combination with a class of acaricides orinsecticides known as insect growth regulators (IGRs). Compoundsbelonging to this group are well known to the practitioner and representa wide range of different chemical classes. These compounds all act byinterfering with the development or growth of the insect pests. Insectgrowth regulators are described, for example, in U.S. Pat. Nos.3,748,356, 3,818,047, 4,225,598, 4,798,837, 4,751,225, EP 0 179 022 orU.K. 2 140 010 as well as U.S. Pat. Nos. 6,096,329 and 6,685,954 (allincorporated herein by reference).

In one embodiment the IGR is a compound that mimics juvenile hormone.Examples of juvenile hormone mimics include azadirachtin, diofenolan,fenoxycarb, hydroprene, kinoprene, methoprene, pyriproxyfen,tetrahydroazadirachtin and4-chloro-2(2-chloro-2-methyl-propyl)-5-(6-iodo-3-pyridylmethoxy)pyridizine-3(2H)-oneExamples of IGRs suitable for use include but are not limited tomethoprene, pyriproxyfen, hydroprene, cyromazine, fluazuron, lufenuron,novaluron, pyrethroids,1-(2,6-difluorobenzoyl)-3-(2-fluoro-4-(trifluoromethyl)phenylurea, andnovaluron.

In one embodiment, the compositions of the invention comprise an indolecompound of formula (I) in combination with methoprene and apharmaceutically acceptable carrier.

In another embodiment, the IGR compound is a chitin synthesis inhibitor.Chitin synthesis inhibitors include chlorofluazuron, cyromazine,diflubenzuron, fluazuron, flucycloxuron, flufenoxuron, hexaflumoron,lufenuron, tebufenozide, teflubenzuron, triflumoron,1-(2,6-difluorobenzoyl)-3-(2-fluoro-4-(trifluoromethyl)phenylurea,1-(2,6-difluoro-benzoyl)-3-(2-fluoro-4-(1,1,2,2-tetrafluoroethoxy)-phenylureaand 1-(2,6-difluorobenzoyl)-3-(2-fluoro-4-trifluoromethyl)phenylurea.

In another embodiment, the compositions of the invention may comprise anindole compound of formula (I) in combination with a formamidine activeagent including, but not limited to, amitraz.

In yet another embodiment of the invention, adulticide insecticides andacaricides can also be added to the composition of the invention. Theseinclude pyrethrins (which include cinerin I, cinerin II, jasmolin I,jasmolin II, pyrethrin I, pyrethrin II and mixtures thereof) andpyrethroids including permethrin, deltamethrin, cyfluthrin, flumethrin,cypermethrin and alpha cypermethrin; and carbamates (which include butare not limited to benomyl, carbanolate, carbaryl, carbofuran,meththiocarb, metolcarb, promacyl, propoxur, aldicarb, butocarboxim,oxamyl, thiocarboxime and thiofanox).

In some embodiments, the compositions of the invention may include oneor more antinematodal agents including, but not limited to, activeagents in the benzimidazoles, imidazothiazoles, tetrahydropyrimidines,organophosphates class of compounds. In some embodiments, benzimidazolesincluding, but not limited to, thiabendazole, cambendazole,parbendazole, oxibendazole, mebendazole, flubendazole, fenbendazole,oxfendazole, albendazole, cyclobendazole, febantel, thiophanate and itso,o-dimethyl analogue may be included in the compositions.

In other embodiments, the compositions may include an imidazothiazolecompounds including, but not limited to, tetramisole, levamisole andbutamisole. In still other embodiments, the compositions of theinvention may include tetrahydropyrimidine active agents including, butnot limited to, pyrantel, oxantel, and morantel. Suitableorganophosphate active agents include, but are not limited to,coumaphos, trichlorfon, haloxon, naftalofos and dichlorvos, heptenophos,mevinphos, monocrotophos, TEPP, and tetrachlorvinphos.

In other embodiments, the compositions may include the antinematodalcompounds phenothiazine, piperazine as the neutral compound and invarious salt forms, diethylcarbamazine, phenols such as disophenol,arsenicals such as arsenamide, ethanolamines such as bephenium, theniumclosylate, and methyridine; cyanine dyes including pyrvinium chloride,pyrvinium pamoate and dithiazanine iodide; isothiocyanates includingbitoscanate, suramin sodium, phthalofyne, and various natural productsincluding, but not limited to, hygromycin B, α-santonin and kainic acid.

In other embodiments, the compositions of the invention may includeother antitrematodal agents. Suitable antitrematodal agents include, butare not limited to, the miracils such as miracil D and mirasan;praziquantel, clonazepam and its 3-methyl derivative, oltipraz,lucanthone, hycanthone, oxamniquine, amoscanate, niridazole, nitroxynil,various bisphenol compounds known in the art including hexachlorophene,bithionol, bithionol sulfoxide and menichlopholan; varioussalicylanilide compounds including tribromsalan, oxyclozanide,clioxanide, rafoxanide, brotianide, bromoxanide and closantel;triclabendazole, diamfenetide, clorsulon, hetolin and emetine.

Anticestodal compounds may also be advantageously used in thecompositions of the invention including, but not limited to, arecolinein various salt forms, bunamidine, niclosamide, nitroscanate,paromomycin and paromomycin II.

In another embodiment of the invention, the compositions may include aspinosyn active agent produced by the soil actinomyceteSaccharopolyspora spinosa (see, for example Salgado V. L. and Sparks T.C., “The Spinosyns: Chemistry, Biochemistry, Mode of Action, andResistance,” in Comprehensive Molecular Insect Science, vol. 6, pp.137-173, 2005) or a semi-synthetic spinosoid active agent. The spinosynsare typically referred to as factors or components A, B, C, D, E, F, G,H, J, K, L, M, N, 0, P, Q, R, S, T, U, V, W, or Y, and any of thesecomponents, or a combination thereof, may be used in the compositions ofthe invention. The spinosyn compound may be a 5,6,5-tricylic ringsystem, fused to a 12-membered macro cyclic lactone, a neutral sugar(rhamnose), and an amino sugar (forosamine). These and other naturalspinosyn compounds, including 21-butenyl spinosyn produced bySaccharopolyspora pagona, which may be used in the compositions of theinvention, may be produced via fermentation by conventional techniquesknown in the art. Other spinosyn compounds that may be used in thecompositions of the invention are disclosed in U.S. Pat. Nos. 5,496,931;5,670,364; 5,591,606; 5,571,901; 5,202,242; 5,767,253; 5,840,861;5,670,486; 5,631,155 and 6,001,981, all incorporated by reference hereinin their entirety. The spinosyn compounds may include, but are notlimited to, spinosyn A, spinosyn D, spinosad, spinetoram, orcombinations thereof. Spinosad is a combination of spinosyn A andspinosyn D, and spinetoram is a combination of 3′-ethoxy-5,6-dihydrospinosyn J and 3′-ethoxy spinosyn L.

In yet other embodiments, the compositions of the invention may includeother active agents that are effective against arthropod parasites.Suitable active agents include, but are not limited to, bromocyclen,chlordane, DDT, endosulfan, lindane, methoxychlor, toxaphene, bromophos,bromophos-ethyl, carbophenothion, chlorfenvinphos, chlorpyrifos,crotoxyphos, cythioate, diazinon, dichlorenthion, diemthoate,dioxathion, ethion, famphur, fenitrothion, fenthion, fospirate,iodofenphos, malathion, naled, phosalone, phosmet, phoxim, propetamphos,ronnel, stirofos, allethrin, cyhalothrin, cypermethrin, deltamethrin,fenvalerate, flucythrinate, permethrin, phenothrin, pyrethrins,resmethrin, benzyl benzoate, carbon disulfide, crotamiton,diflubenzuron, diphenylamine, disulfiram, isobornyl thiocyanato acetate,methoprene, monosulfiram, pirenonylbutoxide, rotenone, triphenyltinacetate, triphenyltin hydroxide, deet, dimethyl phthalate, and thecompounds 1,5a,6,9,9a,9b-hexahydro-4a(4H)-dibenzofurancarboxaldehyde(MGK-11),2-(2-ethylhexyl)-3a,4,7,7a-tetrahydro-4,7-methano-1H-isoindole-1,3(2H)dione(MGK-264), dipropyl-2,5-pyridinedicarboxylate (MGK-326) and2-(octylthio)ethanol (MGK-874).

An antiparasitic agent that can be combined with the compounds of theinvention to form a composition can be a biologically active peptide orprotein including, but not limited to, depsipeptides, which act at theneuromuscular junction by stimulating presynaptic receptors belonging tothe secretin receptor family resulting in the paralysis and death ofparasites. In one embodiment of the depsipeptide, the depsipeptide isemodepside (see Willson et al., Parasitology, January 2003, 126(Pt1):79-86).

In one embodiment of the invention, arylpyrazole compounds such asphenylpyrazoles may be included in the veterinary compositions of theinvention. The arylpyrazoles are known in the art and are suitable forcombination with the isoxazoline compounds in the soft chewablecompositions of the invention. Examples of such arylpyrazole compoundsinclude but are not limited to those described in U.S. Pat. Nos.6,001,384; 6,010,710; 6,083,519; 6,096,329; 6,174,540; 6,685,954,6,998,131 and 7,759,381 (all of which are incorporated herein byreference). A particularly preferred arylpyrazole active agent isfipronil.

An insecticidal agent that can be combined with the compounds of theinvention to form a composition can be a substituted pyridylmethylderivative compound such as imidacloprid. Agents of this class aredescribed above, and for example, in U.S. Pat. No. 4,742,060 or in EP 0892 060. It would be well within the skill level of the practitioner todecide which individual compound can be used in the inventiveformulation to treat a particular infection of an insect.

In certain embodiments, an insecticidal agent that can be combined withthe compositions of the invention is a semicarbazone, such asmetaflumizone.

In another embodiment, the compositions of the invention mayadvantageously include one or more isoxazoline compounds known in theart. These active agents are described in WO 2007/079162, WO 2007/075459and US 2009/0133319, WO 2007/070606 and US 2009/0143410, WO 2009/003075,WO 2009/002809, WO 2009/024541, WO 2005/085216 and US 2007/0066617 andWO 2008/122375, all of which are incorporated herein by reference intheir entirety.

In another embodiment of the invention, nodulisporic acid and itsderivatives (a class of known acaricidal, anthelmintic, anti-parasiticand insecticidal agents) may be added to the compositions of theinvention. These compounds are used to treat or prevent infections inhumans and animals and are described, for example, in U.S. Pat. Nos.5,399,582, 5,962,499, 6,221,894 and 6,399,786, all of which are herebyincorporated by reference in their entirety. The compositions mayinclude one or more of the known nodulisporic acid derivatives in theart, including all stereoisomers, such as those described in theliterature cited above.

In another embodiment, anthelmintic compounds of the amino acetonitrileclass (AAD) of compounds such as monepantel (ZOLVIX) and the like may beadded to the compositions of the invention. These compounds aredescribed, for example, in WO 2004/024704; Sager et al., VeterinaryParasitology, 2009, 159, 49-54; Kaminsky et al., Nature vol. 452, 13Mar. 2008, 176-181. The compositions of the invention may also includearyloazol-2-yl cyanoethylamino compounds such as those described in US2008/0312272 to Soll et al., which is incorporated herein in itsentirety, and thioamide derivatives of these compounds, as described inU.S. patent application Ser. No. 12/582,486, filed Oct. 20, 2009, whichis incorporated herein by reference.

The compositions of the invention may also be combined withparaherquamide compounds and derivatives of these compounds, includingderquantel (see Ostlind et al., Research in Veterinary Science, 1990,48, 260-61; and Ostlind et al., Medical and Veterinary Entomology, 1997,11, 407-408). The paraherquamide family of compounds are known class ofcompounds that include a spirodioxepino indole core with activityagainst certain parasites (see Tet. Lett. 1981, 22, 135; J. Antibiotics1990, 43, 1380, and J. Antibiotics 1991, 44, 492). In addition, thestructurally related marcfortine family of compounds, such asmarcfortines A-C, are also known and may be combined with theformulations of the invention (see J. Chem. Soc.—Chem. Comm. 1980, 601and Tet. Lett. 1981, 22, 1977). Further references to the paraherquamidederivatives can be found, for example, in WO 91/09961, WO 92/22555, WO97/03988, WO 01/076370, WO 09/004432, U.S. Pat. No. 5,703,078 and U.S.Pat. No. 5,750,695, all of which are hereby incorporated by reference intheir entirety.

Dosage forms may contain from about 0.5 mg to about 5 g of a combinationof active agents. In one embodiment of the dosage form, the amount ofactive is present in an amount of from about 1 mg to about 500 mg of anactive agent, typically about 25 mg, about 50 mg, about 100 mg, about200 mg, about 300 mg, about 400 mg, about 500 mg, about 600 mg, about800 mg, or about 1000 mg.

Methods of Treatment

In another aspect, the invention is a method for treating helminthinfestation comprising administration of an anthelmintically effectiveamount of a compound according to Formula (I) to an animal in needthereof. In one embodiment, the helminths are trematodes. In anotherembodiment, the helminths are the liver fluke Fasciola hepatica.

In one embodiment of the invention, methods for the treatment orprevention of a parasitic infestation or infection in a domestic animalare provided, which comprise administering an oral, topical orinjectable composition comprising an effective amount of at least oneindole active agent to the animal. The compositions and methods of theinvention are effective against endoparasites, trematodes in particular,of animals and humans.

In one embodiment, the invention provides methods for the treatment andprevention of parasitic infections and infestations of animals (eitherwild or domesticated), including livestock and companion animals such ascats, dogs, horses, sheep, goats, pigs and cattle, with the aim ofridding these hosts of liver flukes commonly encountered by suchanimals.

By “treating” or “treat” or “treatment” is intended the application oradministration of a composition of the invention to an animal that has aparasitic infestation for the eradication of the parasite or thereduction of the number of the parasites infesting the animal undergoingtreatment. It is noted that the compositions of the invention may beused to prevent such a parasitic infestation.

Additional Active Agents

Additional veterinary/pharmaceutical active ingredients may be used inaccordance with all embodiments and aspects detailed above.

In general, the additional active agent is included in the compositionin an amount of between about 0.1 μg and about 1000 mg. More typically,the additional active agent may be included in an amount of about 10 μgto about 500 mg, about 1 mg to about 300 mg, about 10 mg to about 200 mgor about 10 mg to about 100 mg.

In other embodiments of the invention, the additional active agent maybe included in the composition to deliver a dose of about 5 μg/kg toabout 50 mg/kg per weight of the animal. In other embodiments, theadditional active agent may be present in an amount sufficient todeliver a dose of about 0.01 mg/kg to about 30 mg/kg, about 0.1 mg/kg toabout 20 mg/kg, or about 0.1 mg/kg to about 10 mg/kg of weight ofanimal. In other embodiments, the additional active agent may be presentin a dose of about 5 μg/kg to about 200 μg/kg or about 0.1 mg/kg toabout 1 mg/kg of weight of animal. In still another embodiment of theinvention, the additional active agent is included in a dose betweenabout 0.5 mg/kg to about 50 mg/kg.

Optionally, a fragrance may be added to any of the compositions of theinvention. Fragrances which are useful for the invention include but arenot limited to:

-   -   (i) carboxylic acid esters such as octyl acetate, isoamyl        acetate, isopropyl acetate and isobutyl acetate;    -   (ii) fragrant oils such as lavender oil.

The compositions of the invention are made by mixing the appropriateamount of the active agents, pharmaceutically acceptable carrier ordiluent and optionally a crystallization inhibitor, antioxidant,preservative, film former, etc., to form a composition of the invention.In some embodiments the composition can be obtained by following themethod of making these forms described above by the description ofmaking these forms found in general formulation text known to those inthe art, e.g., Remington—The Science and Practice of Pharmacy (21^(st)Edition) (2005), Goodman & Gilman's The Pharmacological Basis ofTherapeutics (11^(th) Edition) (2005) and Ansel's Pharmaceutical DosageForms and Drug Delivery Systems (8^(th) Edition), edited by Allen etal., Lippincott Williams & Wilkins, (2005).

The inventive formulations may contain other inert ingredients such asantioxidants, preservatives, or pH stabilizers. These compounds are wellknown in the formulation art. Antioxidants such as an alpha tocopherol,ascorbic acid, ascorbyl palmitate, fumaric acid, malic acid, sodiumascorbate, sodium metabisulfate, n-propyl gallate, BHA (butylatedhydroxy anisole), BHT (butylated hydroxy toluene) monothioglycerol andthe like, may be added to the present formulation. The antioxidants aregenerally added to the formulation in amounts of from about 0.01 toabout 2.0%, based upon total weight of the formulation, such as about0.05% to about 1.0%.

Preservatives, such as the parabens (methylparaben and/orpropylparaben), are suitably used in the formulation in amounts rangingfrom about 0.01% to about 2.0%, or about 0.05% to about 1.0%. Otherpreservatives include benzalkonium chloride, benzethonium chloride,benzoic acid, benzyl alcohol, bronopol, butylparaben, cetrimide,chlorhexidine, chlorobutanol, chlorocresol, cresol, ethylparaben,imidurea, methylparaben, phenol, phenoxyethanol, phenylethyl alcohol,phenylmercuric acetate, phenylmercuric borate, phenylmercuric nitrate,potassium sorbate, sodium benzoate, sodium propionate, sorbic acid,thimerosal, and the like. Ranges for these compounds include from about0.01% to about 5%.

Compounds which stabilize the pH of the formulation are alsocontemplated. Again, such compounds are well known to a practitioner inthe art as well as how to use these compounds. Buffering systemsinclude, for example, systems selected from the group consisting ofacetic acid/acetate, malic acid/malate, citric acid/citrate, tartaricacid/tartrate, lactic acid/lactate, phosphoric acid/phosphate,glycine/glycimate, tris, glutamic acid/glutamates or sodium carbonate.

The compositions of the invention are administered in parasiticidallyeffective amounts which are suitable to control the parasite in questionto the desired extent, as described below. In each aspect of theinvention, the compounds and compositions of the invention can beapplied against a single pest or combinations thereof.

The compositions of the invention may be administered continuously, fortreatment or prevention of parasitic infections or infestations. In thismanner, the compositions of the invention deliver an effective amount ofthe active compounds to the animal in need thereof to control the targetparasites. By “effective amount” is intended a sufficient amount of acomposition of the invention to eradicate or reduce the number ofparasites infesting the animal.

In some embodiments, an effective amount of the active agent achieves atleast 70% efficacy against the target parasite. In other embodiments, aneffective amount of the active agent achieves at least 80%, or at least90% efficacy against the target pests. In other embodiments, aneffective amount of the active agent will achieve at least 95%, at least98% or 100% efficacy against the target parasites.

Generally, a dose of from about 0.001 to about 100 mg per kg of bodyweight given as a single dose or in divided doses for a period of from 1to 5 days will be satisfactory, but there can be instances where higheror lower dosage ranges are indicated, and such are within the scope ofthis invention. It is well within the routine skill of the practitionerto determine a particular dosing regimen for a specific host andparasite.

Higher amounts may be provided for very prolonged release in or on thebody of the animal. In another treatment embodiment, the amount ofactive agents for animals which are small in size is greater than about0.01 mg/kg, and in another embodiment for the treatment of small-sizedanimals the amount of active agents is between about 0.01 and about 20mg/kg of weight of animal.

The solutions according to the invention may be applied using any meansknown per se, e.g., using an applicator gun or a metering flask,pipette, syringes, roll on, droppers, capsules, foil packages, vials,twist tip containers and other single dose and multi-dose containers.

In another aspect of the invention, a kit for the treatment orprevention of a parasitic infestation in an animal is provided, whichcomprises at least one isoxazoline active agent together with apharmaceutically acceptable carrier and a dispensing device for topicalapplication of the composition. The dispensing device may be a pipette,syringes, roll on, droppers, capsules, foil packages, vials, twist tipcontainers and other single dose and multi-dose containers, whichincludes an effective dose of each active agent in the pharmaceuticallyacceptable carrier or diluent.

An important aspect of the invention is to provide a multiple-usecontainer comprising a topical composition of the invention, from whichaccurate single dose aliquots of the long lasting topical formulationsmay be administered. The formulation must remain stable with repetitiveexposure to the outside environment, particularly oxygen and water. Thisembodiment may be particularly useful with the very long lastingformulations of the invention that require administration to an animalinfrequently, such as once every 3-6 months, or similar. Some solventssuch as ethers (including DMI (dimethyl isosorbide) and the like) giverise to peroxides, which then yield ketones and aldehydes that may befurther degraded to acids. The presence of acids may contribute to thedegradation of acid hydrolysis-susceptible molecules. Thus, formulationstability is particularly important for the multi-dose containerapplication, where the formulations can be exposed to oxygen and waterduring multiple rounds of opening and closing. Importantly, it was foundthat the use of certain antioxidants such as BHT and BHA efficientlyinhibit the degradation of the active agent in ether solvents. Forexample, a 12% (w/v) solution of an indole compound of the invention inDMI exhibited no significant change in assay over the course of aneleven week accelerated stability study at 50° C. in clear glasscontainers.

EXAMPLES

The invention is further described by the following non-limitingexamples which further illustrate the invention and are not intended(nor should they be interpreted to) limit the scope of the invention.

Compound number 332 (i.e.,5-[2-chloro-4-(trifluoromethyl)phenyl]-2-(trifluoromethyl)-1H-indole)was prepared as follows.

Into a 500-mL round-bottom flask, was placed 4-bromo-2-methylaniline (10g, 53.75 mmol, 1.00 equiv), dichloromethane (150 mL), trifluoroacetyl2,2,2-trifluoroacetate (13.6 g, 64.75 mmol, 1.20 equiv). The resultingsolution was stirred for 1 h at 30° C. The resulting mixture wasconcentrated under vacuum. This resulted in 15 g (89%) ofN-(4-bromo-2-methylphenyl)-2,2,2-trifluoroacetamide as a white solid.

Next, into a 50-mL round-bottom flask, was placedN-(4-bromo-2-methylphenyl)-2,2,2-trifluoroacetamide (100 mg, 0.35 mmol,1.00 equiv), CCl₄ (3 mL), NBS (63 mg, 0.35 mmol, 1.00 equiv). Theresulting solution was heated to reflux for 6 h under a 100 wattincandescent bulb and a infrared bub. The resulting mixture wasconcentrated under vacuum. The residue was applied onto a silica gelcolumn with PE/EA (petroleum ether/ethyl acetate, 8:1). This resulted in80 mg (56%) ofN-[4-bromo-2-(bromomethyl)phenyl]-2,2,2-trifluoroacetamide as a whitesolid.

¹H NMR (400 MHz, CDCl₃) δ: 7.83 (d, J=8.4 Hz, 1H), 7.55-˜7.62 (m, 2H),4.44 (s, 3H).

Next, into a 100-mL round-bottom flask, was placedN-[4-bromo-2-(bromomethyl)phenyl]-2,2,2-trifluoroacetamide (600 mg, 1.66mmol, 1.00 equiv), tol (10 mL), PPh₃ (437 mg, 1.67 mmol, 1.00 equiv).The resulting solution was stirred for 3 h at 70° C. The solids werecollected by filtration. This resulted in 600 mg (52%) of[[5-bromo-2-(trifluoroacetamido)phenyl]methyl]triphenylphosphoniumbromide as a white solid.

Next, into a 50-mL round-bottom flask, was placed[[5-bromo-2-(trifluoroacetamido)phenyl]methyl]triphenylphosphoniumbromide (400 mg, 0.64 mmol, 1.00 equiv), N,N-dimethylformamide (10 mL),TEA (triethylamine, 324 mg, 3.20 mmol, 5.00 equiv). The resultingsolution was stirred for 2 h at 130° C. The reaction was then quenchedby the addition of 10 ml of water. The resulting solution was extractedwith 3×10 mL of ethyl acetate and the organic layers combined. Theresulting mixture was washed with 3×20 mL of brine. The mixture wasdried over anhydrous sodium sulfate and concentrated under vacuum. Thisresulted in 300 mg (crude) of 5-bromo-2-(trifluoromethyl)-1H-indole asbrown oil.

Finally, into a 50-mL round-bottom flask purged and maintained with aninert atmosphere of nitrogen, was placed5-bromo-2-(trifluoromethyl)-1H-indole (100 mg, 0.38 mmol, 1.00 equiv),dioxane (5 mL), [2-chloro-4-(trifluoromethyl)phenyl]boronic acid (169mg, 0.75 mmol, 1.99 equiv), Pd(PPh₃)₄ (43 mg, 0.04 mmol, 0.10 equiv),K₃PO₄ (240 mg, 1.13 mmol, 2.99 equiv), water (0.5 mL). The resultingsolution was stirred overnight at 100° C. The reaction was then quenchedby the addition of 10 ml of water. The resulting solution was extractedwith 3×10 mL of ethyl acetate and the organic layers combined. Theresulting mixture was washed with 3×10 ml, of brine. The resultingmixture was concentrated under vacuum. The residue was applied onto aTLC plate with PE/EA (10:1). This resulted in 26.6 mg (17%) of5-[2-chloro-4-(trifluoromethyl)phenyl]-2-(trifluoromethyl)-1H-indole aslight yellow oil. (ES, m/z): [M−H]⁻ 362; ¹H NMR (400 MHz, CDCl₃) δ: 7.80(s, 1H), 7.72 (s, 1H), 7.53-˜7.68 (m, 3H), 7.47 (d, J=6.0 Hz, 1H), 6.95(s, 1H).

Compound number 334 (i.e.,6-[2-chloro-4-(trifluoromethyl)phenyl]-2-(trifluoromethyl)-1H-indole)was prepared as follows.

First, into a 250-mL round-bottom flask, was placed4-bromo-1-methyl-2-nitrobenzene (5 g, 23.14 mmol, 1.00 equiv), ethylacetate (100 mL), SnCl₂.H₂O (20 g). The resulting solution was stirredovernight at 30° C. The pH value of the solution was adjusted to 10 withsodium hydroxide (5 mol/L). The resulting solution was extracted with3×30 mL of ethyl acetate and the organic layers combined. The resultingmixture was washed with 3×50 mL of brine. The organic phase was driedover anhydrous sodium sulfate and concentrated under vacuum. Thisresulted in 4 g (93%) of 5-bromo-2-methylaniline as brown oil.

Next, into a 100-mL round-bottom flask, was placed5-bromo-2-methylaniline (2 g, 10.75 mmol, 1.00 equiv), dichloromethane(40 mL), trifluoroacetyl 2,2,2-trifluoroacetate (2.7 g, 12.86 mmol, 1.20equiv). The resulting solution was stirred for 1 h at 0° C. Theresulting mixture was concentrated under vacuum. This resulted in 2.3 g(76%) of N-(5-bromo-2-methylphenyl)-2,2,2-trifluoroacetamide as a lightbrown solid.

Next, into a 100-mL round-bottom flask, was placedN-(5-bromo-2-methylphenyl)-2,2,2-trifluoroacetamide (500 mg, 1.77 mmol,1.00 equiv), CCl₄ (10 mL), NBS (315 mg, 1.77 mmol, 1.00 equiv). Theresulting solution was heated to reflux for 4 h under a 100 wattincandescent bulb. The solids were filtered out. The resulting mixturewas concentrated under vacuum. This resulted in 600 mg (crude) ofN-[5-bromo-2-(bromomethyl)phenyl]-2,2,2-trifluoroacetamide as a lightyellow solid.

Next, into a 100-mL round-bottom flask, was placedN-[5-bromo-2-(bromomethyl)phenyl]-2,2,2-trifluoroacetamide (600 mg, 1.66mmol, 1.00 equiv), tol (10 mL), PPh₃ (435 mg, 1.66 mmol, 1.00 equiv).The resulting solution was stirred for 4 h at 70° C. The solids werecollected by filtration and washed with EA 10 ml*2. This resulted in 700mg (64%) of[[4-bromo-2-(trifluoroacetamido)phenyl]methyl]triphenylphosphoniumbromide as a white solid.

Next, into a 50-mL round-bottom flask, was placed[[4-bromo-2-(trifluoroacetamido)phenyl]-methyl]triphenylphosphoniumbromide (350 mg, 0.56 mmol, 1.00 equiv), N,N-dimethylformamide (10 mL),TEA (567 mg, 5.60 mmol, 9.98 equiv). The resulting solution was stirredfor 2 h at 120° C. The reaction was then quenched by the addition of 10ml of water. The resulting solution was extracted with 3×10 mL of ethylacetate and the organic layers combined. The resulting mixture waswashed with 3×20 mL of brine. The organic phase was dried over anhydroussodium sulfate and concentrated under vacuum. This resulted in 120 mg(73%) of 6-bromo-2-(trifluoromethyl)-1H-indole as brown oil.

Finally, into a 50-mL round-bottom flask purged and maintained with aninert atmosphere of nitrogen, was placed6-bromo-2-(trifluoromethyl)-1H-indole (80 mg, 0.30 mmol, 1.00 equiv),dioxane (5 mL), [2-chloro-4-(trifluoromethyl)phenyl]boronic acid (135mg, 0.60 mmol, 1.99 equiv), Pd(PPh₃)₄ (34.9 mg, 0.03 mmol, 0.10 equiv),K₃PO₄ (192 mg, 0.90 mmol, 2.99 equiv), water (0.5 mL). The resultingsolution was stirred overnight at 100° C. The reaction was then quenchedby the addition of 10 ml of water. The resulting solution was extractedwith 3×10 mL of ethyl acetate and the organic layers combined. Theresulting mixture was washed with 3×10 mL of brine. The resultingmixture was concentrated under vacuum. The residue was applied onto aTLC plate with PE/EA (10/1). This resulted in 113 mg (92%) of6-[2-chloro-4-(trifluoromethyl)phenyl]-2-(trifluoromethyl)-1H-indole asyellow oil. (ES, m/z): [M−H]⁻ 362; ¹H NMR (400 MHz, CDCl₃) δ: 7.84 (s,1H), 7.63-7.77 (m, 3H), 7.55 (s, 1H), 7.22 (d, J=7.5 Hz, 1H), 6.97 (s,1H).

Compound number 336 (i.e.,6-chloro-5-[2-chloro-4-(trifluoromethyl)phenyl]-2-(trifluoromethyl)-1H-indole)was made as follows.

First, into a 500-mL 3-necked round-bottom flask purged and maintainedwith an inert atmosphere of nitrogen, was placed dichloromethane (400mL), 4-bromo-5-chloro-2-methylaniline (25 g, 113.38 mmol, 1.00 equiv).This was followed by the addition of TFAA (trifluoroacetic anhydride,27.4 g, 130.46 mmol, 1.15 equiv) dropwise with stirring at 0° C. in awater/ice bath. The resulting solution was allowed to react, withstirring, for an additional 1 h at room temperature. The resultingmixture was concentrated under vacuum. This resulted in 30 g (84%) ofN-(4-bromo-5-chloro-2-methylphenyl)-2,2,2-trifluoroacetamide as a whitesolid.

Next, into a 500-mL round-bottom flask purged and maintained with aninert atmosphere of nitrogen, was placed CCl₄ (300 mL),N-(4-bromo-5-chloro-2-methylphenyl)-2,2,2-trifluoroacetamide (15 g,47.39 mmol, 1.00 equiv), NBS (8 g, 44.95 mmol, 0.95 equiv). Theresulting solution was stirred for 4 h at 80° C. with infrared light andinitiated by light. This reaction was repeated for 1 times. Then thereaction mixture was cooled. The solids were filtered out and washedwith enough CCl₄. The filtrate was concentrated under vacuum. Thisresulted in 40 g (crude) ofN-[4-bromo-2-(bromomethyl)-5-chlorophenyl]-2,2,2-trifluoroacetamide as abrown solid.

Next, into a 1-L round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed toluene (800 mL),N-[4-bromo-2-(bromomethyl)-5-chlorophenyl]-2,2,2-trifluoroacetamide (40g, 101.16 mmol, 1.00 equiv), PPh₃ (29.5 g, 112.47 mmol, 1.20 equiv). Theresulting solution was stirred for 2 h at 80° C. in an oil bath. Thereaction mixture was cooled.

The solids were collected by filtration, and washed with enough EA. Thenthe solid was dried under infrared light. This resulted in 45 g (68%) of[[5-bromo-4-chloro-2-(trifluoroacetamido)phenyl]methyl]triphenylphosphoniumbromide as a white solid.

Next, into a 1-L round-bottom flask, was placed N,N-dimethylformamide(600 mL),[[5-bromo-4-chloro-2-(trifluoroacetamido)phenyl]methyl]triphenylphosphoniumbromide (45 g, 68.42 mmol, 1.00 equiv), TEA (34 g, 336.00 mmol, 5.00equiv). The resulting solution was stirred for 2 h at 120° C. in an oilbath. The reaction mixture was cooled. The reaction mixture was pouredinto 1.5 L of water/ice. The resulting solution was extracted with 3×700mL of ethyl acetate and the organic layers combined. The resultingmixture was washed with 3×600 mL of brine. The organic phase wasconcentrated under vacuum. The residue was applied onto a silica gelcolumn with ethyl acetate/petroleum ether (1:10). This resulted in 17 g(83%) of 5-bromo-6-chloro-2-(trifluoromethyl)-1H-indole as a whitesolid.

Finally, into a 250-mL 3-necked round-bottom flask purged and maintainedwith an inert atmosphere of nitrogen, was placed dioxane (120 mL), water(20 mL), 5-bromo-6-chloro-2-(trifluoromethyl)-1H-indole (6 g, 20.10mmol, 1.00 equiv), [2-chloro-4-(trifluoromethyl)phenyl]boronic acid (9g, 40.11 mmol, 2.00 equiv), K₃PO₄ (12.7 g, 59.83 mmol, 3.00 equiv),Pd(PPh₃)₄ (1.85 g, 1.60 mmol, 0.08 equiv). The resulting solution wasstirred overnight at 100° C. in an oil bath. This reaction was repeatedfor 1 times. The reaction mixture was cooled. The resulting solution wasdiluted with 300 mL of ethyl acetate. The resulting mixture was washedwith 3×80 mL of brine. The resulting mixture was concentrated undervacuum. The residue was applied onto a silica gel column withdichloromethane. This resulted in 11.0 g (69%) of6-chloro-5-[2-chloro-4-(trifluoromethyl)phenyl]-2-(trifluoromethyl)-1H-indoleas a white solid. LC-MS-PH-MRL-209-336-0: (ES, m/z): 396[M−H]⁻H-NMR-PH-MRL-209-336-0: (300 MHz, DMSO, ppm): δ 12.65 (s, 1H), 8.02 (s,1H), 7.83 (d, J=9.0 Hz, 1H), 7.71 (d, J=9.0 Hz, 2H), 7.62˜7.65 (d, J=9.0Hz, 1H), 7.14 (s, 1H).

Compound number 337 (i.e.,6-chloro-5-[2-chloro-4-(trifluoromethyl)phenyl]-2-methyl-1H-indole) wasmade as follows.

First, into a 500-mL 3-necked round-bottom flask purged and maintainedwith an inert atmosphere of nitrogen, was placed methanol/DCM (150/100mL), 2-amino-4-chlorobenzoic acid (10 g, 58.28 mmol, 1.00 equiv). Thiswas followed by the addition of TMSCHN₂ (30.7 mL, 1.05 equiv) dropwisewith stirring at 0° C. The resulting solution was stirred overnight atroom temperature. The resulting mixture was concentrated under vacuum.The resulting mixture was washed with 1×20 mL of n-hexane. This resultedin 10.1 g (93%) of methyl 2-amino-4-chlorobenzoate as a yellow solid.

Next, into a 250-mL 3-necked round-bottom flask, was placedN,N-dimethylformamide (200 mL), methyl 2-amino-4-chlorobenzoate (8 g,43.10 mmol, 1.00 equiv), NBS (7.7 g, 1.00 equiv). The resulting solutionwas stirred for 1 h at room temperature. The resulting solution wasdiluted with 600 mL of ice/water. The resulting solution was extractedwith 3×200 mL of ethyl acetate and the organic layers combined. Theresulting mixture was washed with 3×150 mL of Brine. The resultingorganic phase was concentrated under vacuum. The solid was washed with 4mL/20 mL of ether/hexane. The solid was dried in an oven under reducedpressure. This resulted in 8.0 g (70%) of methyl2-amino-5-bromo-4-chlorobenzoate as a pink solid.

Next, into a 250-mL 3-necked round-bottom flask purged and maintainedwith an inert atmosphere of nitrogen, was placed tetrahydrofuran (200mL). This was followed by the addition of LiAlH₄ (1.26 g, 37.14 mmol,1.10 equiv), in portions. To this was added methyl2-amino-5-bromo-4-chlorobenzoate (8 g, 30.25 mmol, 1.00 equiv) dropwisewith stirring at −78° C. The resulting solution was stirred for 1 h atroom temperature. The reaction was then quenched by the addition of 200mL of water. The resulting solution was extracted with 3×150 mL of ethylacetate and the organic layers combined. The resulting mixture waswashed with 3×100 mL of brine. The organic phase was dried overanhydrous sodium sulfate and concentrated under vacuum. This resulted in7 g (98%) of (2-amino-5-bromo-4-chlorophenyl)methanol as a light yellowsolid.

Next, into a 50-mL 3-necked round-bottom flask purged and maintainedwith an inert atmosphere of nitrogen, was placed tetrahydrofuran (30mL), (2-amino-5-bromo-4-chlorophenyl)methanol (1.27 g, 5.37 mmol, 1.00equiv), TEA (1.35 g, 13.34 mmol, 2.50 equiv). This was followed by theaddition of acetyl chloride (880 mg, 11.21 mmol, 2.10 equiv) dropwisewith stirring at 0° C. The resulting solution was stirred for 0.5 h atroom temperature. And then the reaction mixture was prepared for thenext step.

Next, added the 10% LiPH/H₂O (10 mL) to step four reaction mixture. Theresulting solution was stirred for 0.5 h at room temperature. Theresulting solution was diluted with 30 mL of water and extracted with3×20 mL of ethyl acetate and the organic layers combined. The resultingmixture was washed with 3×15 mL of Brine. The organic phase was driedover anhydrous sodium sulfate and concentrated under vacuum. Thisresulted in 1.3 g (83%) ofN-[4-bromo-5-chloro-2-(hydroxymethyl)phenyl]acetamide as a white solid.

Next, into a 50-mL 3-necked round-bottom flask purged and maintainedwith an inert atmosphere of nitrogen, was placed dichloromethane (30 g,353.23 mmol, 75.68 equiv),N-[4-bromo-5-chloro-2-(hydroxymethyl)phenyl]acetamide (1.3 g, 4.67 mmol,1.00 equiv). This was followed by the addition of tribromophosphane (490mg, 1.81 mmol, 0.40 equiv) dropwise with stirring at 5° C. The resultingsolution was stirred for 0.5 h at 5° C. The temperature was increased toroom temperature naturally. The resulting solution was diluted with 50mL of DCM. The reaction was then quenched by the addition of 1 mL ofwater. The resulting mixture was washed with 3×10 mL of Brine. Theorganic phase was dried over anhydrous sodium sulfate and concentratedunder vacuum. This resulted in 1.3 g (63%) ofN-[4-bromo-2-(bromomethyl)-5-chlorophenyl]acetamide as a white solid.

Next, into a 50-mL round-bottom flask purged and maintained with aninert atmosphere of nitrogen, was placed toluene (30 mL),N-[4-bromo-2-(bromomethyl)-5-chlorophenyl]acetamide (1.3 g, 3.81 mmol,1.00 equiv), triphenylphosphine (1.1 g, 4.19 mmol, 1.10 equiv). Theresulting solution was stirred for 2 h at 70° C. in an oil bath. Thereaction mixture was cooled to room temperature. The solids werecollected by filtration and washed with enough EtOAc. The solid wasdried in an oven under reduced pressure. This resulted in 1.0 g (44%) of[(5-bromo-4-chloro-2-acetamidophenyl)methyl]triphenylphosphonium bromideas a white solid.

Next, into a 50-mL round-bottom flask purged and maintained with aninert atmosphere of nitrogen, was placed toluene (30 mL),[(5-bromo-4-chloro-2-acetamidophenyl)methyl]triphenylphosphanium bromide(1.0 g, 1.66 mmol, 1.00 equiv). The reaction mixture was stirred at 120°C. in an oil bath, then added (tert-butoxy)potassium (740 mg, 6.59 mmol,4.00 equiv) in two portions for 1.5 hours. The reaction mixture wascooled to room temperature. The resulting mixture was concentrated undervacuum. The residue was applied onto a silica gel column with ethylacetate/petroleum ether (1:25-1:10). This resulted in 100 mg (25%) of5-bromo-6-chloro-2-methyl-1H-indole as a white solid.

Finally, into a 100-mL round-bottom flask purged and maintained with aninert atmosphere of nitrogen, was placed dioxane (10 mL), water (2 mL),5-bromo-6-chloro-2-methyl-1H-indole (100 mg, 0.41 mmol, 1.00 equiv),[2-chloro-4-(trifluoromethyl)phenyl]boronic acid (184 mg, 0.82 mmol,2.00 equiv), K₃PO₄ (260 mg, 1.22 mmol, 3.00 equiv), Pd(PPh₃)₄ (47 mg,0.04 mmol, 0.10 equiv). The resulting solution was stirred for 3 h at100° C. in an oil bath. The reaction mixture was cooled to roomtemperature. The resulting solution was diluted with 40 mL of ethylacetate. The resulting mixture was washed with 3×10 mL of Brine. Theorganic phase was dried over anhydrous sodium sulfate and concentratedunder vacuum. The residue was purified by thin layer chromatographydeveloped with ethyl acetate/PE (1:5) This resulted in 22.9 mg (16%) of6-chloro-5-[2-chloro-4-(trifluoromethyl)phenyl]-2-methyl-1H-indole as aoff-white solid. (ES, m/z): 342[M−H]⁻

H-NMR-PH-MRL-209-337-0: (300 MHz, DMSO, ppm): δ 11.23 (s, 1H), 7.97˜8.00(m, 1H), 7.78 (dd, J=1.2, 8.1 Hz, 1H), 7.59 (d, J=7.8 Hz, 1H), 7.45 (s,1H), 7.35 (s, 1H), 6.18˜6.19 (m, 1H), 2.40 (s, 3H).

Compound number 338 (i.e.,6-chloro-5-[2-chloro-4-(trifluoromethyl)phenyl]-2-ethyl-1H-indole) wasmade as follows.

First, into a 100-mL 3-necked round-bottom flask purged and maintainedwith an inert atmosphere of nitrogen, was placed tetrahydrofuran (50mL), (2-amino-5-bromo-4-chlorophenyl)methanol (2 g, 8.46 mmol, 1.00equiv), TEA (1.88 g, 18.58 mmol, 2.20 equiv). This was followed by theaddition of propanoyl chloride (1.64 g, 17.73 mmol, 2.00 equiv) dropwisewith stirring at 5° C. The resulting solution was stirred for 0.5 h atroom temperature. Then the reaction mixture was prepared for the nextstep.

Next, 15% LiOH/H₂O (40 mL) was added to the step one reaction mixture.The resulting solution was stirred for 0.5 h at room temperature. Theresulting solution was diluted with 100 mL of water and extracted with3×100 mL of ethyl acetate and the organic layers combined. The resultingmixture was washed with 3×80 mL of Brine. The organic phase was driedover anhydrous sodium sulfate and concentrated under vacuum. Thisresulted in 2.5 g (99%) ofN-[4-bromo-5-chloro-2-(hydroxymethyl)phenyl]-propanamide as a lightyellow solid.

Next, into a 100-mL 3-necked round-bottom flask purged and maintainedwith an inert atmosphere of nitrogen, was placed dichloromethane (40mL), N-[4-bromo-5-chloro-2-(hydroxymethyl)phenyl]propanamide (1.6 g,5.47 mmol, 1.00 equiv). This was followed by the addition of PBr₃ (570mg, 2.11 mmol, 0.40 equiv) dropwise with stirring at 5° C. The resultingsolution was stirred for 0.5 h at 5° C. The resulting solution wasdiluted with 50 mL of DCM. The reaction was then quenched by theaddition of 1 mL of water. The resulting mixture was washed with 3×20 mLof Brine. The organic phase was dried over anhydrous sodium sulfate andconcentrated under vacuum. This resulted in 1.4 g (72%) ofN-[4-bromo-2-(bromomethyl)-5-chlorophenyl]propanamide as a light yellowsolid.

Next, into a 50-mL round-bottom flask purged and maintained with aninert atmosphere of nitrogen, was placed toluene (30 mL),N-[4-bromo-2-(bromomethyl)-5-chlorophenyl]propanamide (1.4 g, 3.94 mmol,1.00 equiv), triphenylphosphine (1.13 g, 4.31 mmol, 1.10 equiv). Theresulting solution was stirred for 2 h at 70° C. in an oil bath. Thereaction mixture was cooled to room temperature. The solids werecollected by filtration, and the solid was washed with enough EtOAc. Thesolid was dried in an oven under reduced pressure. This resulted in 1.05g (43%) of[(5-bromo-4-chloro-2-propanamidophenyl)methyl]triphenylphosphoniumbromide as a white solid.

Next, into a 50-mL round-bottom flask purged and maintained with aninert atmosphere of nitrogen, was placed toluene (20 mL),[(5-bromo-4-chloro-2-propanamidophenyl)methyl]-triphenylphosphoniumbromide (950 mg, 1.54 mmol, 1.00 equiv). The reaction mixture wasstirred at 120° C., then (tert-butoxy)potassium (689 mg, 6.14 mmol, 4.00equiv), in two portions for 1.5 hours. The reaction mixture was cooledto room temperature. The resulting mixture was concentrated undervacuum. The residue was applied onto a silica gel column with ethylacetate/petroleum ether (1:25-1:15). This resulted in 90 mg (23%) of5-bromo-6-chloro-2-ethyl-1H-indole as a light yellow solid.

Finally, into a 50-mL round-bottom flask purged and maintained with aninert atmosphere of nitrogen, was placed dioxane (10 mL), water (2 mL),5-bromo-6-chloro-2-ethyl-1H-indole (70 mg, 0.27 mmol, 1.00 equiv),[2-chloro-4-(trifluoromethyl)phenyl]boronic acid (122 mg, 0.54 mmol,2.00 equiv), K3PO4 (172 mg, 0.81 mmol, 3.00 equiv), Pd(PPh3)4 (31 mg,0.03 mmol, 0.10 equiv). The resulting solution was stirred for 3 h at100° C. in an oil bath. The reaction mixture was cooled. The resultingsolution was diluted with 40 mL of ethyl acetate. The resulting mixturewas washed with 3×10 mL of Brine. The mixture was dried over anhydroussodium sulfate and concentrated under vacuum. The residue was dissolvedin 5 mL of methanol. The crude product was purified by Prep-HPLC. Thisresulted in 10.8 mg (11%) of6-chloro-5-[2-chloro-4-(trifluoromethyl)phenyl]-2-ethyl-1H-indole as alight brown solid. (ES, m/z): 356 [M−H]⁻

H-NMR-PH-MRL-209-338-0: (300 MHz, DMSO, ppm): δ 11.25 (s, 1H), 8.00 (s,1H), 7.79 (d, J=8.1 Hz, 1H), 7.59 (d, J=7.2 Hz, 1H), 7.45 (s, 1H), 7.37(s, 1H), 6.21 (s, 1H), 2.72˜2.77 (m, 2H), 1.28 (t, J=7.5 Hz, 3H).

Compound number 339 (i.e.,6-chloro-5-[2-chloro-4-(trifluoromethyl)phenyl]-2-propyl-1H-indole) wasmade as follows.

First, into a 100-mL 3-necked round-bottom flask purged and maintainedwith an inert atmosphere of nitrogen, was placed tetrahydrofuran (60mL), (2-amino-5-bromo-4-chlorophenyl)methanol (1.84 g, 7.78 mmol, 1.00equiv), TEA (1.74 g, 17.20 mmol, 2.20 equiv). This was followed by theaddition of butanoyl chloride (1.73 g, 16.24 mmol, 2.10 equiv) dropwisewith stirring at 5° C. The resulting solution was stirred for 0.5 h atroom temperature.

Then the reaction mixture was prepared for the next step.

Next, 10% LiOH/H₂O (40 mL) was added to the step one reaction mixture.The resulting solution was stirred for 0.5 hour at room temperature. Theresulting solution was diluted with 100 mL of water. The resultingsolution was extracted with 3×100 mL of ethyl acetate and the organiclayers combined. The resulting mixture was washed with 3×80 mL of Brine.The organic phase was dried over anhydrous sodium sulfate andconcentrated under vacuum. This resulted in 2.6 g (91%) ofN-[4-bromo-5-chloro-2-(hydroxymethyl)phenyl]butanamide as a light yellowsolid.

Next, into a 100-mL 3-necked round-bottom flask purged and maintainedwith an inert atmosphere of nitrogen, was placed dichloromethane (40mL), N-[4-bromo-5-chloro-2-(hydroxymethyl)phenyl]butanamide (1.5 g, 4.89mmol, 1.00 equiv). This was followed by the addition of PBr₃ (520 mg,1.92 mmol, 0.40 equiv) dropwise with stirring at 5° C. The resultingsolution was stirred for 0.5 h at 5° C. The resulting solution wasdiluted with 50 mL of DCM. The reaction was then quenched by theaddition of 1 mL of water. The resulting mixture was washed with 3×20 mLof Brine. The mixture was dried over anhydrous sodium sulfate andconcentrated under vacuum. This resulted in 1.3 g (72%) ofN-[4-bromo-2-(bromomethyl)-5-chlorophenyl]-butanamide as a white solid.

Next, into a 50-mL round-bottom flask purged and maintained with aninert atmosphere of nitrogen, was placed toluene (30 mL),N-[4-bromo-2-(bromomethyl)-5-chlorophenyl]butanamide (1 g, 2.71 mmol,1.00 equiv), triphenylphosphine (780 mg, 2.97 mmol, 1.10 equiv). Theresulting solution was stirred for 2 h at 70° C. in an oil bath. Thereaction mixture was cooled to room temperature. The solids werecollected by filtration, and the solid was washed with enough EtOAc.This resulted in 1.14 g (67%) of[(5-bromo-2-butanamido-4-chlorophenyl)methyl]-triphenylphosphoniumbromide as a white solid.

Next, into a 50-mL round-bottom flask purged and maintained with aninert atmosphere of nitrogen, was placed toluene (20 mL),[(5-bromo-2-butanamido-4-chlorophenyl)methyl]-triphenylphosphoniumbromide (1.1 g, 1.74 mmol, 1.00 equiv), the reaction mixture was stirredat 120° C. in an oil bath, then added (tert-butoxy)potassium (780 mg,6.95 mmol, 4.00 equiv) in two portions for 1.5 hours. The reactionmixture was cooled. The resulting mixture was concentrated under vacuum.The residue was applied onto a silica gel column with ethylacetate/petroleum ether (1:25-1:20). This resulted in 100 mg (21%) of5-bromo-6-chloro-2-propyl-1H-indole as a white solid.

Finally, into a 50-mL round-bottom flask purged and maintained with aninert atmosphere of nitrogen, was placed dioxane (10 mL), water (2 mL),5-bromo-6-chloro-2-propyl-1H-indole (70 mg, 0.26 mmol, 1.00 equiv),[2-chloro-4-(trifluoromethyl)phenyl]-boronic acid (115 mg, 0.51 mmol,2.00 equiv), K₃PO₄ (163 mg, 0.77 mmol, 3.00 equiv), Pd(PPh₃)₄ (30 mg,0.03 mmol, 0.10 equiv). The resulting solution was stirred for 3 h at100° C. in an oil bath. The reaction mixture was cooled to roomtemperature. The resulting solution was diluted with 40 mL of ethylacetate. The resulting mixture was washed with 3×10 mL of Brine. Theorganic phase was dried over anhydrous sodium sulfate and concentratedunder vacuum. The residue was dissolved in 5 mL of methanol. The crudeproduct was purified by Prep-HPLC. This resulted in 16 mg (17%) of6-chloro-5-[2-chloro-4-(trifluoromethyl)-phenyl]-2-propyl-1H-indole ascolorless oil. (ES, m/z): 370 [M−H]⁻

H-NMR-PH-MRL-209-339-0: (300 MHz, DMSO, ppm): δ 11.23 (s, 1H), 7.98 (s,1H), 7.78˜7.80 (m, 1H), 7.59 (d, J=7.5 Hz, 1H), 7.45 (s, 1H), 7.37 (s,1H), 6.22 (s, 1H), 2.70 (t, J=7.5 Hz, 2H), 1.68˜1.75 (m, 2H), 0.95 (t,J=7.2 Hz, 3H).

Compound number 340 (i.e.,6-chloro-5-[2-chloro-4-(trifluoromethyl)phenyl]-2-(pentafluoroethyl)-1H-indole)was made as follows.

First, into a 100-mL 3-necked round-bottom flask, was placed4-bromo-5-chloro-2-methylaniline (600 mg, 2.72 mmol, 1.00 equiv),dichloromethane (18 mL), pentafluoropropanoyl2,2,3,3,3-pentafluoropropanoate (846 mg, 2.73 mmol, 1.00 equiv). Theresulting solution was stirred for 30 min at 0° C. in a water/ice bath.The resulting mixture was concentrated under vacuum. This resulted in740 mg (74%) ofN-(4-bromo-5-chloro-2-methylphenyl)-2,2,3,3,3-pentafluoropropanamide asa light brown solid.

Next, into a 100-mL round-bottom flask, was placedN-(4-bromo-5-chloro-2-methylphenyl)-2,2,3,3,3-pentafluoropropanamide(400 mg, 1.09 mmol, 1.00 equiv), CCl₄ (10 mL), NBS (195.1 mg, 1.10 mmol,1.00 equiv). The resulting solution was refluxed for 2 h under a 100 wincandescent bulb. The solids were filtered out. The resulting mixturewas concentrated under vacuum. This resulted in 480 mg (99%) ofN-[4-bromo-2-(bromomethyl)-5-chlorophenyl]-2,2,3,3,3-pentafluoropropanamideas a yellow solid.

Next, into a 100-mL round-bottom flask, was placedN-[4-bromo-2-(bromomethyl)-5-chlorophenyl]-2,2,3,3,3-pentafluoropropanamide(470 mg, 1.06 mmol, 1.00 equiv), tooulene (10 mL), PPh₃ (334.4 mg, 1.27mmol, 1.20 equiv). The resulting solution was stirred for 2 h at 70° C.in an oil bath. The resulting mixture was concentrated under vacuum.This resulted in 620 mg (83%) of[[5-bromo-4-chloro-2-(pentafluoropropanamido)phenyl]methyl]triphenylphosphoniumbromide as a yellow solid.

Next, into a 100-mL round-bottom flask, was placed[[5-bromo-4-chloro-2-(pentafluoropropanamido)phenyl]methyl]triphenylphosphoniumbromide (450 mg, 0.64 mmol, 1.00 equiv), N,N-dimethylformamide (15 mL),TEA (321 mg, 3.17 mmol, 5.00 equiv). The resulting solution was stirredfor 2 h at 120° C. in an oil bath. The resulting solution was dilutedwith 20 mL of H₂O. The resulting solution was extracted with 3×20 mL ofethyl acetate and the organic layers combined. The resulting mixture waswashed with 3×20 mL of brine. The organic phase was dried over anhydroussodium sulfate and concentrated under vacuum. This resulted in 160 mg(72%) of 5-bromo-6-chloro-2-(pentafluoroethyl)-1H-indole as brown oil.

Finally, into a 50-mL round-bottom flask purged and maintained with aninert atmosphere of nitrogen, was placed5-bromo-6-chloro-2-(pentafluoroethyl)-1H-indole (100 mg, 0.29 mmol, 1.00equiv), dioxane (6 mL), water (1 mL), K₃PO₄ (183.3 mg, 0.86 mmol, 3.00equiv), Pd(PPh₃)₄ (33.3 mg, 0.03 mmol, 0.10 equiv),[2-chloro-4-(trifluoromethyl)phenyl]boronic acid (130 mg, 0.58 mmol,2.00 equiv). The resulting solution was stirred overnight at 100° C. inan oil bath. The resulting mixture was concentrated under vacuum. Theresidue was applied onto a silica gel column with ethylacetate/petroleum ether (1:4). This resulted in 33 mg (26%) of6-chloro-5-[2-chloro-4-(trifluoromethyl)phenyl]-2-(pentafluoroethyl)-1H-indoleas a white solid. (ES, m/z): [M−H]⁻ 446; ¹H NMR (CDCl₃, 300 MHz) δ: 8.50(s, 1H), 7.77 (s, 1H), 7.71-7.57 (m, 3H), 7.43 (d, J=9.0 Hz, 1H), 6.99(s, 1H).

Compound number 341 (i.e.,6-chloro-5-[2-chloro-4-(trifluoromethyl)phenyl]-2-(heptafluoropropyl)-1H-indole)was made as follows.

First, into a 50-mL 3-necked round-bottom flask, was placed4-bromo-5-chloro-2-methylcyclohexan-1-amine (500 mg, 2.21 mmol, 1.00equiv), dichloromethane (10 mL), heptafluorobutanoyl2,2,3,3,4,4,4-heptafluorobutanoate (931.8 mg, 2.27 mmol, 1.00 equiv).The resulting solution was stirred for 30 min at 0° C. in a water/icebath. The resulting mixture was concentrated under vacuum. This resultedin 640 mg (70%) ofN-(4-bromo-5-chloro-2-methylphenyl)-2,2,3,3,4,4,4-heptafluorobutanamideas a brown solid.

Next, into a 50-mL round-bottom flask, was placedN-(4-bromo-5-chloro-2-methylphenyl)-2,2,3,3,4,4,4-heptafluorobutanamide(400 mg, 0.96 mmol, 1.00 equiv), CCl₄ (10 mL), NBS (172 mg, 0.97 mmol,1.00 equiv). The resulting solution was refluxed for 2 h under a 100 wincandescent bulb. The resulting mixture was concentrated under vacuum.This resulted in 520 mg (crude) ofN-[4-bromo-2-(bromomethyl)-5-chlorophenyl]-2,2,3,3,4,4,4-heptafluorobutanamideas a brown solid.

Next, into a 50-mL round-bottom flask, was placedN-[4-bromo-2-(bromomethyl)-5-chlorophenyl]-2,2,3,3,4,4,4-heptafluorobutanamide(300 mg, 0.61 mmol, 1.00 equiv), toulene (6 mL), PPh₃ (219.07 mg, 0.84mmol, 1.20 equiv). The resulting solution was stirred for 2 h at 75° C.in an oil bath. The solids were collected by filtration. The filter cakewas washed with 3×5 mL of EA. This resulted in 400 mg (87%) of[[5-bromo-4-chloro-2-(heptafluorobutanamido)phenyl]methyl]triphenylphosphoniumbromide as a pink solid.

Next, into a 50-mL round-bottom flask, was placed[[5-bromo-4-chloro-2-(heptafluorobutanamido)phenyl]methyl]triphenylphosphoniumbromide (250 mg, 0.33 mmol, 1.00 equiv), N,N-dimethylformamide (8 mL),TEA (166.6 mg, 1.65 mmol, 5.00 equiv). The resulting solution wasstirred for 2 h at 120° C. The resulting solution was diluted with 10 mLof H₂O. The resulting solution was extracted with 3×15 mL of ethylacetate and the organic layers combined. The resulting mixture waswashed with 3×15 mL of brine. The mixture was dried over anhydroussodium sulfate and concentrated under vacuum. This resulted in 120 mg(91%) of 5-bromo-6-chloro-2-(heptafluoropropyl)-1H-indole as brown oil.

Finally, into a 50-mL round-bottom flask, was placed5-bromo-6-chloro-2-(heptafluoropropyl)-1H-indole (100 mg, 0.25 mmol,1.00 equiv), dioxane (6 mL), water (1 mL), K₃PO₄ (160.2 mg, 0.75 mmol,3.00 equiv), [2-chloro-4-(trifluoromethyl)phenyl]boronic acid (113.4 mg,0.51 mmol, 2.00 equiv), Pd(PPh₃)₄ (29.1 mg, 0.03 mmol, 0.10 equiv). Theresulting solution was stirred overnight at 100° C. in an oil bath. Theresulting mixture was concentrated under vacuum. The residue was appliedonto a silica gel column with ethyl acetate/petroleum ether (1:1). Thisresulted in 29.1 mg (23%) of6-chloro-5-[2-chloro-4-(trifluoromethyl)phenyl]-2-(heptafluoropropyl)-1H-indoleas a white solid. (ES, m/z): [M−H]⁻ 496; ¹H NMR (CDCl₃, 300 MHz) δ: 8.49(s, 1H), 7.77 (s, 1H), 7.63-7.57 (m, 3H), 7.45-7.43 (m, 1H), 6.99 (s,1H).

Compound number 342 (i.e.,6-chloro-5-[4-chloro-2-(trifluoromethyl)phenyl]-2-(trifluoromethyl)-1H-indole)was made as follows.

Into a 50-mL round-bottom flask, was placed a solution of5-bromo-6-chloro-2-(trifluoromethyl)-1H-indole (80 mg, 0.27 mmol, 1.00equiv) in dioxane/H₂O (1.8:0.3 mL),[4-chloro-2-(trifluoromethyl)phenyl]boronic acid (120 mg, 0.53 mmol,2.00 equiv), Pd(PPh₃)₄ (31 mg, 0.03 mmol, 0.10 equiv), K₃PO₄ (171 mg,3.00 equiv). The resulting solution was stirred overnight at 100° C. inan oil bath under N2 protection. The resulting mixture was concentratedunder vacuum. The residue was applied onto a silica gel column withethyl acetate/hexane (1:10). This resulted in 26.2 mg (25%) of6-chloro-5-[4-chloro-2-(trifluoromethyl)phenyl]-2-(trifluoromethyl)-1H-indoleas a white solid. (ES, m/z): [M−H]⁻ 396.

¹H NMR (CDCl₃, 400 MHz) δ: 8.44 (s, 1H), 7.76-7.77 (m, 1H), 7.54-7.60(m, 3H), 7.28 (s, 1H), 6.90-6.93 (m, 1H).

Compound number 343 (i.e.,6-chloro-5-(4-chlorophenyl)-2-(trifluoromethyl)-1H-indole) was made asfollows.

Into a 40-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed dioxane (10 mL), water (2 mL),5-bromo-6-chloro-2-(trifluoromethyl)-1H-indole (80 mg, 0.27 mmol, 1.00equiv), (4-chlorophenyl)boronic acid (84 mg, 0.54 mmol, 2.00 equiv),K3PO4 (170 mg, 0.80 mmol, 3.00 equiv), Pd(PPh3)4 (31 mg, 0.03 mmol, 0.10equiv). The resulting solution was stirred overnight at 100° C. in anoil bath. The reaction mixture was cooled. The resulting solution wasdiluted with 50 mL of ethyl acetate. The resulting mixture was washedwith 3×20 mL of brine. The resulting mixture was concentrated undervacuum. The residue was purified by preparative TLC (EtOAc:PE=1:20).This resulted in 14.1 mg (16%) of6-chloro-5-(4-chlorophenyl)-2-(trifluoromethyl)-1H-indole as a whitesolid. (ES, m/z): 328[M−H]⁻

H-NMR-PH-MRL-209-343-0: (300 MHz, DMSO, ppm): δ 12.55 (s, 1H), 7.72 (s,1H), 7.65 (s, 1H), 7.51˜7.54 (m, 2H), 7.44˜7.47 (m, 2H), 7.10 (s, 1H).

Compound number 344 (i.e.,6-chloro-5-(4-fluorophenyl)-2-(trifluoromethyl)-1H-indole) was made asfollows.

Into a 40-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed dioxane (10 mL), water (2 mL),5-bromo-6-chloro-2-(trifluoromethyl)-1H-indole (80 mg, 0.27 mmol, 1.00equiv), (4-fluorophenyl)boronic acid (75 mg, 0.54 mmol, 2.00 equiv),K₃PO₄ (170 mg, 0.80 mmol, 3.00 equiv), Pd(PPh₃)₄ (31 mg, 0.03 mmol, 0.10equiv). The resulting solution was stirred overnight at 100° C. in anoil bath. The reaction mixture was cooled. The resulting solution wasdiluted with 50 mL of ethyl acetate. The resulting mixture was washedwith 3×20 mL of brine. The organic phase was concentrated under vacuum.The residue was purified by preparative TLC (EtOAc:PE=1:20). Thisresulted in 41.5 mg (49%) of6-chloro-5-(4-fluorophenyl)-2-(trifluoromethyl)-1H-indole as a whitesolid. (ES, m/z): 312[M−H]⁻

H-NMR-PH-MRL-209-344-0: (300 MHz, DMSO, ppm): δ 12.51 (s, 1H), 7.70 (s,1H), 7.64 (s, 1H), 7.45˜7.49 (m, 2H), 7.29 (t, J=9.0 Hz, 2H), 7.09 (s,1H).

Compound number 345 (i.e.,6-chloro-2-(trifluoromethyl)-5-[4-(trifluoromethyl)phenyl]-1H-indole)was made as follows.

Into a 40-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed dioxane (10 mL), water (2 mL),5-bromo-6-chloro-2-(trifluoromethyl)-1H-indole (80 mg, 0.27 mmol, 1.00equiv), [4-(trifluoromethyl)phenyl]boronic acid (101 mg, 0.53 mmol, 2.00equiv), K₃PO4 (170 mg, 0.80 mmol, 3.00 equiv), Pd(PPh₃)₄ (31 mg, 0.03mmol, 0.10 equiv). The resulting solution was stirred overnight at 100°C. in an oil bath. The reaction mixture was cooled. The resultingsolution was diluted with 50 mL of ethyl acetate. The resulting mixturewas washed with 3×20 mL of brine. The resulting mixture was concentratedunder vacuum. The residue was purified by preparative TLC(EtOAc:PE=1:20). This resulted in 67 mg (69%) of6-chloro-2-(trifluoromethyl)-5-[4-(trifluoromethyl)phenyl]-1H-indole asa white solid. (ES, m/z): 362[M−H]⁻

H-NMR-PH-MRL-209-345-0: (300 MHz, DMSO, ppm): δ 12.59 (s, 1H), 7.81˜7.84(m, 2H), 7.77 (s, 1H), 7.66˜7.69 (m, 3H), 7.12 (s, 1H).

Compound number 346 (i.e.,6-chloro-5-(2-chlorophenyl)-2-(trifluoromethyl)-1H-indole) was made asfollows.

Into a 50-mL round-bottom flask, was placed a solution of5-bromo-6-chloro-2-(trifluoromethyl)-1H-indole (80 mg, 0.27 mmol, 1.00equiv) in dioxane/H₂O (1.8:0.3 mL), (2-chlorophenyl)boronic acid (84 mg,0.54 mmol, 2.00 equiv), Pd(PPh₃)₄ (31 mg, 0.03 mmol, 0.10 equiv), K₃PO₄(171 mg, 3.00 equiv). The resulting solution was stirred overnight at100° C. in an oil bath. The resulting mixture was concentrated undervacuum. The residue was applied onto a silica gel column with ethylacetate/hexane (1:10). This resulted in 20 mg (23%) of6-chloro-5-(2-chlorophenyl)-2-(trifluoromethyl)-1H-indole as colorlessoil. (ES, m/z): [M−H]⁻ 328. ¹H NMR (CDCl₃, 400 MHz) δ: 8.45 (s, 1H),7.59-7.60 (m, 2H), 7.53-7.49 (s, 1H), 7.30-7.40 (m, 3H), 6.96 (s, 1H).

Compound number 347 (i.e.,6-chloro-5-(3-chlorophenyl)-2-(trifluoromethyl)-1H-indole) was made asfollows.

Into a 50-mL round-bottom flask, was placed a solution of5-bromo-6-chloro-2-(trifluoromethyl)-1H-indole (80 mg, 0.27 mmol, 1.00equiv) in dioxane/H₂O (1.8:0.3 mL), (3-chlorophenyl)boronic acid (84 mg,0.54 mmol, 2.00 equiv), Pd(PPh₃)₄ (31 mg, 0.03 mmol, 0.10 equiv), K₃PO₄(171 mg, 3.00 equiv). The resulting solution was stirred overnight at100° C. in an oil bath under N₂ protection. The solid was dried in anoven under reduced pressure. The residue was applied onto a silica gelcolumn with ethyl acetate/hexane (1:10). This resulted in 12.6 mg (14%)of 6-chloro-5-(3-chlorophenyl)-2-(trifluoromethyl)-1H-indole as a lightbrown solid. (ES, m/z): [M−H]⁻ 328. ¹H NMR (CDCl₃, 400 MHz) δ: 8.44 (s,1H), 7.56-7.61 (m, 2H), 7.44 (s, 1H), 7.35 (s, 3H), 6.92 (s, 1H).

Compound number 348 (i.e.,6-chloro-5-(3,4-dichlorophenyl)-2-(trifluoromethyl)-1H-indole) was madeas follows.

Into a 50-mL round-bottom flask, was placed a solution of5-bromo-6-chloro-2-(trifluoromethyl)-1H-indole (80 mg, 0.27 mmol, 1.00equiv) in dioxane/H₂O (1.8:0.3 mL), (3,4-dichlorophenyl)boronic acid(102 mg, 0.53 mmol, 2.00 equiv), Pd(PPh₃)₄ (31 mg, 0.03 mmol, 0.10equiv), K₃PO₄ (171 mg, 3.00 equiv). The resulting solution was stirredovernight at 100° C. in an oil bath under N₂ protection. The resultingmixture was concentrated under vacuum. The residue was applied onto asilica gel column with ethyl acetate/hexane (1:10). This resulted in38.7 mg (40%) of6-chloro-5-(3,4-dichlorophenyl)-2-(trifluoromethyl)-1H-indole as a whitesolid. (ES, m/z): [M−H]⁻ 362. ¹H NMR (CDCl₃, 400 MHz) δ: 8.42 (s, 1H),7.49-7.60 (m, 4H), 7.28-7.31 (m, 1H), 6.93 (s, 1H).

Compound number 349 (i.e.,6-chloro-5-(pyridin-4-yl)-2-(trifluoromethyl)-1H-indole) was made asfollows.

Into a 50-mL round-bottom flask, was placed a solution of5-bromo-6-chloro-2-(trifluoromethyl)-1H-indole (80 mg, 0.27 mmol, 1.00equiv) in dioxane/H₂O (1.8:0.3 mL), (pyridin-4-yl)boronic acid (66 mg,0.54 mmol, 2.00 equiv), Pd(PPh₃)₄ (31 mg, 0.03 mmol, 0.10 equiv), K₃PO₄(171 mg, 0.81 mmol, 3.00 equiv). The resulting solution was stirredovernight at 100° C. in an oil bath under N₂ protection. The resultingmixture was concentrated under vacuum. The residue was applied onto asilica gel column with ethyl acetate/hexane (1:10). This resulted in24.2 mg (30%) of 6-chloro-5-(pyridin-4-yl)-2-(trifluoromethyl)-1H-indoleas a white solid. (ES, m/z): [M−H]⁻ 295. ¹H NMR (CDCl₃, 400 MHz) δ:12.62 (s, 1H), 8.64-8.66 (m, 2H), 7.79 (s, 1H), 7.68 (s, 1H), 7.48-7.50(m, 2H), 7.12 (s, 1H).

Compound number 350 (i.e.,6-chloro-5-(4-chlorophenoxy)-2-(trifluoromethyl)-1H-indole was made asfollows.

First, into a 80-mL round-bottom flask, was placed1,2-dichloro-4-methyl-5-nitrobenzene (1 g, 4.9 mmol, 1.00 equiv),N,N-dimethylformamide (20 mL), potassium carbonate (2 g, 14.5 mmol, 3.00equiv), 4-chlorophenol (0.62 g, 4.8 mmol, 1.00 equiv). The resultingsolution was stirred for 2 h at 120° C. in an oil bath. The resultingsolution was diluted with 20 mL of H₂O. The resulting solution wasextracted with 3×20 mL of ethyl acetate and the organic layers combined.The resulting mixture was washed with 3×20 mL of brine. The organicphase was dried over anhydrous sodium sulfate and concentrated undervacuum. This resulted in 1.2 g (78%) of1-chloro-2-(4-chlorophenoxy)-4-methyl-5-nitrobenzene as a black solid.

Next, into a 100-mL round-bottom flask, was placed methanol (40 mL),Raney Ni (200 mg), 1-chloro-2-(4-chlorophenoxy)-4-methyl-5-nitrobenzene(1.1 g, 3.69 mmol, 1.00 equiv). The flask was evacuated and flushedthree times with hydrogen. The mixture was stirred 1 h at roomtemperature under an atmosphere of hydrogen. The solids were filteredout. The resulting mixture was concentrated under vacuum. This resultedin 800 mg (81%) of 5-chloro-4-(4-chlorophenoxy)-2-methylaniline as ablack solid.

Next, into a 100-mL 3-necked round-bottom flask purged and maintainedwith an inert atmosphere of nitrogen, was placed dichloromethane (40mL), 5-chloro-4-(4-chlorophenoxy)-2-methylaniline (800 mg, 2.98 mmol,1.00 equiv). This was followed by the addition of TFAA (752 mg, 3.58mmol, 1.20 equiv) dropwise with stirring at 0° C. The resulting solutionwas stirred for 1 h at room temperature. The resulting mixture wasconcentrated under vacuum. This resulted in 600 mg (55%) ofN-[5-chloro-4-(4-chlorophenoxy)-2-methylphenyl]-2,2,2-trifluoroacetamideas a red solid.

Next, into a 50-mL round-bottom flask purged and maintained with aninert atmosphere of nitrogen, was placed tetrachloromethane (20 mL),N-[5-chloro-4-(4-chlorophenoxy)-2-methylphenyl]-2,2,2-trifluoroacetamide(600 mg, 1.65 mmol, 1.00 equiv), boranylidene(sulfanyl)amine (288 mg,4.89 mmol, 0.98 equiv). The resulting solution was stirred for 3 h at80° C. by the infrared light and initiated by light. The reactionmixture was cooled. The solids were filtered out, and washed with enoughCCl₄. Then the filtrate was concentrated under vacuum. This resulted in500 mg (crude) ofN-[2-(bromomethyl)-5-chloro-4-(4-chlorophenoxyl)phenyl]-2,2,2-trifluoroacetamideas a brown solid.

Next, into a 50-mL round-bottom flask purged and maintained with aninert atmosphere of nitrogen, was placed toluene (30 mL),N-[2-(bromomethyl)-5-chloro-4-(4-chlorophenoxyl)phenyl]-2,2,2-trifluoroacetamide(500 mg, 1.13 mmol, 1.00 equiv), triphenylphosphine (354 mg, 1.35 mmol,1.20 equiv). The resulting solution was stirred for 2 h at 80° C. in anoil bath. The reaction mixture was cooled. The solids were filtered outand washed with enough EtOAc. The solid was dried under infrared light.This resulted in 400 mg (50%) ofN-[2-[(bromotriphenyl-^[5]-phosphanyl)methyl]-5-chloro-4-(4-chlorophenoxyl)phenyl]-2,2,2-trifluoroacetamideas a white solid.

Finally, into a 40-mL round-bottom flask, was placedN,N-dimethylformamide (10 mL),[[4-chloro-5-(4-chlorophenoxy)-2-(trifluoroacetamido)phenyl]methyl]triphenylphosphoniumbromide (150 mg, 0.21 mmol, 1.00 equiv), TEA (107 mg, 1.06 mmol, 5.00equiv). The resulting solution was stirred for 2 h at 120° C. in an oilbath. The reaction mixture was cooled. The resulting solution was pouredinto 50 mL of water/ice. The resulting solution was extracted with 3×30mL of ethyl acetate and the organic layers combined. The resultingmixture was washed with 3×15 mL of brine. The resulting mixture wasconcentrated under vacuum. The residue was purified by preparative TLC(EtOAc:PE=1:20). This resulted in 41 mg (56%) of6-chloro-5-(4-chlorophenoxy)-2-(trifluoromethyl)-1H-indole as a whitesolid. (ES, m/z): 344[M−H]⁻

H-NMR-PH-MRL-209-350-0: (300 MHz, DMSO, ppm): δ 12.56 (s, 1H), 7.69 (s,1H), 7.57 (s, 1H), 7.36˜7.41 (m, 2H), 7.05 (s, 1H), 6.88˜6.93 (m, 2H).

Compound number 351 (i.e.,6-chloro-5-(2-chlorophenoxy)-2-(trifluoromethyl)-1H-indole) was made asfollows.

First, into a 100-mL round-bottom flask, was placed1,2-dichloro-4-methyl-5-nitrobenzene (1 g, 4.85 mmol, 1.00 equiv),N,N-dimethylformamide (20 g, 273.64 mmol, 56.38 equiv), potassiumcarbonate (2 g, 14.47 mmol, 3.00 equiv), 2-chlorophenol (621 mg, 4.83mmol, 1.00 equiv). The resulting solution was stirred for 1 h at 100° C.in an oil bath. The resulting solution was diluted with 20 mL of H₂O.The resulting solution was extracted with 3×20 mL of ethyl acetate andthe organic layers combined. The resulting mixture was washed with 3×20mL of brine. The mixture was dried over anhydrous sodium sulfate andconcentrated under vacuum. This resulted in 1.24 g (91%) of1-chloro-2-(2-chlorophenyl)-4-methyl-5-nitrobenzene as a brown solid.

Next, into a 100-mL round-bottom flask, was placed methanol (30 mL),Raney Ni (200 mg, 20% W/W equiv),1-chloro-2-(2-chlorophenoxy)-4-methyl-5-nitrobenzene (1 g, 3.35 mmol,1.00 equiv), hydrogen (g, enough). The resulting solution was stirredfor 1 h at room temperature. The solids were filtered out. The resultingmixture was concentrated under vacuum. This resulted in 700 mg (78%) of5-chloro-4-(2-chlorophenoxy)-2-methylaniline as a brown solid.

Next, into a 100-mL 3-necked round-bottom flask, was placeddichloromethane (30 mL), 5-chloro-4-(2-chlorophenoxy)-2-methylaniline(700 mg, 2.61 mmol, 1.00 equiv), This was followed by the addition ofTFAA (658 mg, 3.13 mmol, 1.20 equiv) dropwise with stirring at 0° C. Theresulting solution was stirred for 1 h at room temperature. Theresulting mixture was concentrated under vacuum. This resulted in 700 mg(74%) ofN-[5-chloro-4-(2-chlorophenoxy)-2-methylphenyl]-2,2,2-trifluoroacetamideas a brown solid.

Next, into a 50-mL round-bottom flask purged and maintained with aninert atmosphere of nitrogen, was placed tetrachloromethane (30 mL), NBS(335 mg, 1.88 mmol, 0.98 equiv),N-[5-chloro-4-(2-chlorophenoxy)-2-methylphenyl]-2,2,2-trifluoroacetamide(700 mg, 1.92 mmol, 1.00 equiv). The resulting solution was stirred for4 h at 80° C. by infrared light and initiated by light. The reactionmixture was cooled to room temperature. The solids were filtered out andwashed with enough CCl₄. The filtrate was concentrated under vacuum.This resulted in 500 mg (crude) ofN-[2-(bromomethyl)-5-chloro-4-(2-chlorophenoxyl)phenyl]-2,2,2-trifluoroacetamideas a light brown solid. The product was used in the next step directlywithout further purification.

Next, into a 50-mL round-bottom flask purged and maintained with aninert atmosphere of nitrogen, was placed toluene (30 mL),N-[2-(bromomethyl)-5-chloro-4-(2-chlorophenoxyl)phenyl]-2,2,2-trifluoroacetamide(500 mg, 1.13 mmol, 1.00 equiv), triphenylphosphine (603 mg, 2.30 mmol,1.20 equiv). The resulting solution was stirred for 2 h at 80° C. in anoil bath. The reaction mixture was cooled to room temperature. Thesolids were collected by filtration. The solid was dried in an ovenunder reduced pressure. This resulted in 320 mg (40%) of[[4-chloro-5-(2-chlorophenoxy)-2-(trifluoroacetamido)phenyl]methyl]triphenylphosphoniumbromide as a white solid.

Finally, into a 40-mL round-bottom flask purged and maintained with aninert atmosphere of nitrogen, was placed N,N-dimethylformamide (10 mL),[[4-chloro-5-(2-chlorophenoxy)-2-(trifluoroacetamido)phenyl]methyl]triphenylphosphoniumbromide (150 mg, 0.21 mmol, 1.00 equiv), TEA (107 mg, 1.06 mmol, 5.00equiv). The resulting solution was stirred for 2 h at 120° C. in an oilbath. The reaction mixture was cooled to room temperature. The resultingsolution was diluted with 1×50 mL of H₂O. The resulting solution wasextracted with 3×30 mL of ethyl acetate and the organic layers combined.The resulting mixture was washed with 3×15 ml, of brine. The resultingorganic phase was concentrated under vacuum. The residue was purified byprep-HPLC. This resulted in 51 mg (69%) of6-chloro-5-(2-chlorophenoxy)-2-(trifluoromethyl)-1H-indole as a lightbrown solid. (ES, m/z): 344[M−H]⁻

H-NMR-PH-MRL-209-351-0: (300 MHz, DMSO, ppm): δ 12.51 (brs, 1H), 7.70(s, 1H), 7.59 (dd, J=1.5, 9.0 Hz, 1H), 7.44 (s, 1H), 7.28 (dt, J=3.0,9.0 Hz, 1H), 7.14 (dt, J=1.5, 7.8 Hz, 1H), 7.03 (s, 1H), 6.78 (dd,J=1.5, 8.4 Hz, 1H).

Compound number 352 (i.e.,6-chloro-2-(trifluoromethyl)-5-[4-(trifluoromethyl)phenoxy]-1H-indole)was made as follows.

First, into a 100-mL round-bottom flask, was placed1,2-dichloro-4-methyl-5-nitrobenzene (1 g, 4.85 mmol, 1.00 equiv),N,N-dimethylformamide (20 mL), potassium carbonate (2 g, 14.47 mmol,3.00 equiv), 4-(trifluoromethyl)phenol (786 mg, 4.85 mmol, 1.00 equiv).The resulting solution was stirred for 2 h at 100° C. in an oil bath.The resulting solution was diluted with 20 mL of H₂O. The resultingsolution was extracted with 3×20 mL of ethyl acetate and the organiclayers combined. The resulting mixture was washed with 3×20 mL of brine.The organic phase was dried over anhydrous sodium sulfate andconcentrated under vacuum. This resulted in 1.2 g (78%) of1-chloro-4-methyl-5-nitro-2-[4-(trifluoromethyl)phenyl]benzene as abrown solid.

Next, into a 100-mL round-bottom flask, was placed methanol (30 mL),Raney Ni (200 mg),1-chloro-4-methyl-5-nitro-2-[4-(trifluoromethyl)phenoxy]benzene (1.1 g,3.32 mmol, 1.00 equiv). The flask was evacuated and flushed three timeswith hydrogen. The mixture was stirred 1 h at room temperature under anatmosphere of hydrogen. The solids were filtered out. The resultingmixture was concentrated under vacuum. This resulted in 800 mg (80%) of5-chloro-2-methyl-4-[4-(trifluoromethyl)phenoxy]aniline as a brownsolid.

Next, into a 100-mL 3-necked round-bottom flask purged and maintainedwith an inert atmosphere of nitrogen, was placed dichloromethane (30mL), 5-chloro-2-methyl-4-[4-(trifluoromethyl)phenoxy]aniline (800 mg,2.65 mmol, 1.00 equiv). This was followed by the addition of TFAA (670mg, 3.19 mmol, 1.20 equiv) dropwise with stirring at 0° C. The resultingsolution was stirred for 1 h at room temperature. The resulting mixturewas concentrated under vacuum. This resulted in 800 mg (76%) ofN-[5-chloro-2-methyl-4-[4-(trifluoromethyl)phenoxy]phenyl]-2,2,2-trifluoroacetamideas a brown solid.

Next, into a 50-mL round-bottom flask purged and maintained with aninert atmosphere of nitrogen, was placed tetrachloromethane (30 mL),N-[5-chloro-2-methyl-4-[4-(trifluoromethyl)phenoxy]phenyl]-2,2,2-trifluoroacetamide(800 mg, 2.01 mmol, 1.00 equiv), NBS (430 mg, 7.30 mmol, 0.98 equiv).The resulting solution was stirred for 2 days at 80° C. by infraredlight and initiated by light. The reaction mixture was cooled. Thesolids were filtered out and washed with enough CCl₄. The filtrate wasconcentrated under vacuum. This resulted in 600 mg (crude) ofN-[2-(bromomethyl)-5-chloro-4-[4-(trifluoromethyl)phenoxy]phenyl]-2,2,2-trifluoroacetamideas a light brown solid.

Next, into a 50-mL round-bottom flask purged and maintained with aninert atmosphere of nitrogen, was placed toluene (30 mL),N-[2-(bromomethyl)-5-chloro-4-[4-(trifluoromethyl)phenoxy]phenyl]-2,2,2-trifluoroacetamide(600 mg, 1.26 mmol, 1.00 equiv), triphenylphosphine (628 mg, 2.39 mmol,1.20 equiv). The resulting solution was stirred for 2 h at 80° C. in anoil bath. The reaction mixture was cooled to room temperature. Thesolids were collected by filtration and washed with EtOAc. The solid wasdried under infrared light. This resulted in 700 mg (75%) of[[4-chloro-2-(trifluoroacetamido)-5-[4-(trifluoromethyl)phenoxy]phenyl]methyl]triphenylphosphoniumbromide as a white solid.

Finally, into a 40-mL round-bottom flask, was placedN,N-dimethylformamide (10 mL),[[4-chloro-2-(trifluoroacetamido)-5-[4-(trifluoromethyl)phenoxy]phenyl]methyl]triphenylphosphaniumbromide (150 mg, 0.20 mmol, 1.00 equiv), TEA (102 mg, 1.01 mmol, 5.00equiv). The resulting solution was stirred for 2 h at 120° C. in an oilbath. The reaction mixture was cooled to room temperature. The resultingsolution was poured into 50 mL of water/ice. The resulting solution wasextracted with 3×30 mL of ethyl acetate and the organic layers combined.The solvent was removed under reduced pressure. The residue was purifiedby preparative TLC (EtOAc:PE=20:1). This resulted in 30.8 mg (40%) of6-chloro-2-(trifluoromethyl)-5-[4-(trifluoromethyl)phenoxy]-1H-indole asa white solid. (ES, m/z): 378[M−H]⁻

H-NMR-PH-MRL-209-352-0: (300 MHz, DMSO, ppm): δ 12.63 (s, 1H), 7.69˜7.73(m, 4H), 7.02˜7.08 (m, 3H).

Compound number 354 (i.e.,6-chloro-5-(pyridin-4-yl)-2-(trifluoromethyl)-1H-indole hydrochloride)was made as follows.

First, a solution of 4-bromo-5-chloro-2-methylaniline (5 g, 22.68 mmol,1.00 equiv), TFAA (5.5 g, 26.19 mmol, 1.50 equiv) in dichloromethane (50mL) was placed into a 100-mL round-bottom flask. The resulting solutionwas stirred for 4 h at room temperature. The resulting mixture wasconcentrated under vacuum. This resulted in 7.9 g (crude) ofN-(4-bromo-5-chloro-2-methylphenyl)-2,2,2-trifluoroacetamide as a brownsolid.

Next, a solution ofN-(4-bromo-5-chloro-2-methylphenyl)-2,2,2-trifluoroacetamide (7.9 g,24.96 mmol, 1.00 equiv) in CCl₄ (100 mL), NBS (4.38 g, 24.61 mmol, 0.98equiv) was placed into a 250-mL round-bottom flask. The resultingsolution was stirred for 3 h at 60° C. The resulting mixture wasconcentrated under vacuum. The residue was applied onto a silica gelcolumn with ethyl acetate/petroleum ether (1:10). This resulted in 10 g(crude) ofN-[4-bromo-2-(bromomethyl)-5-chlorophenyl]-2,2,2-trifluoroacetamide as abrown solid.

Next, a solution ofN-[4-bromo-2-(bromomethyl)-5-chlorophenyl]-2,2,2-trifluoroacetamide (11g, 27.82 mmol, 1.00 equiv), PPh₃ (8.1 g, 30.88 mmol, 1.10 equiv) in Tol(100 mL) was placed into a 250-mL round-bottom flask. The resultingsolution was stirred overnight at 100° C. The resulting mixture wasconcentrated under vacuum and washed by MeOH. This resulted in 9.2 g(50%) of[[5-bromo-4-chloro-2-(trifluoroacetamido)phenyl]methyl]triphenylphosphoniumbromide as a yellow solid.

Next, a solution of[[5-bromo-4-chloro-2-(trifluoroacetamido)phenyl]methyl]triphenylphosphoniumbromide (9.2 g, 13.99 mmol, 1.00 equiv), triethylamine (7.1 g, 70.17mmol, 5.00 equiv) in N,N-dimethylformamide (120 mL) was placed into a250-mL round-bottom flask. The resulting solution was stirred for 5 h atroom temperature. The reaction was then quenched by the addition of 50mL of water. The resulting solution was extracted with 3×100 mL of ethylacetate and the organic layers combined and concentrated under vacuum.The residue was applied onto a silica gel column with ethylacetate/petroleum ether (1:20). This resulted in 4.8 g (crude) of5-bromo-6-chloro-2-(trifluoromethyl)-1H-indole as a yellow solid.

Finally, a solution of 5-bromo-6-chloro-2-(trifluoromethyl)-1H-indole(100 mg, 0.34 mmol, 1.00 equiv), K₃PO₄ (214.33 mg, 1.01 mmol, 3.00equiv), Pd(PPh₃)₄ (39 mg, 0.03 mmol, 0.10 equiv), (pyridin-4-yl)boronicacid (82.23 mg, 0.67 mmol, 2.00 equiv) in dioxane/H₂O (6 mL/1 mL) wasplaced into a 50-mL round-bottom flask purged and maintained with aninert atmosphere of nitrogen. The resulting solution was stirredovernight at 100° C. The crude product was purified by Prep-HPLC. Thisresulted in 31 mg (28%) of6-chloro-5-(pyridin-4-yl)-2-(trifluoromethyl)-1H-indole hydrochloride asa off-white solid. (ES, m/z): [M+H]⁺ 333; ¹H NMR (300 MHz, DMSO):12.89-12.84 (m, 1H), 8.92 (d, J=6.6 Hz, 2H), 8.18-8.04 (m, 2H), 7.96 (d,J=4.2 Hz, 1H), 7.78 (d, J=3.3 Hz, 1H), 7.20 (s, 1H).

Compound number 355 (i.e.,3-chloro-4-[6-chloro-2-(trifluoromethyl)-1H-indol-5-yl]benzonitrile) wasmade as follows.

Into a 100-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed dioxane (60 mL),5-bromo-6-chloro-2-(trifluoromethyl)-1H-indole (3 g, 10.05 mmol, 1.00equiv), Pin₂B₂ (5.1 g, 2.00 equiv), potassium acetate (2.95 g, 30.06mmol, 3.00 equiv), Pd(dppf)Cl₂ (500 mg, 0.68 mmol, 0.07 equiv). Theresulting solution was stirred overnight at 80° C. in an oil bath. Thereaction mixture was cooled. The resulting solution was diluted with 50mL of H₂O. The resulting solution was extracted with 3×50 mL of ethylacetate and the organic layers combined. The resulting mixture waswashed with 3×30 mL of brine. The organic phase was dried over anhydroussodium sulfate and concentrated under vacuum. The residue was appliedonto a silica gel column with ethyl acetate/petroleum ether (1:30-1:15).This resulted in 2.5 g (72%) of6-chloro-5-(tetramethyl-1,3,2-dioxaborolan-2-yl)-2-(trifluoromethyl)-1H-indoleas a white solid.

Next, into a 50-mL round-bottom flask purged and maintained with aninert atmosphere of nitrogen, was placed dioxane (5 mL), water (1 mL),6-chloro-5-(tetramethyl-1,3,2-dioxaborolan-2-yl)-2-(trifluoromethyl)-1H-indole(100 mg, 0.29 mmol, 1.00 equiv), 4-bromo-3-chlorobenzonitrile (62.6 mg,0.29 mmol, 1.00 equiv), (phosphoperoxy)potassium; dipotassium (184 mg,0.866 mmol, 3.00 equiv), Pd(dppf)Cl₂.CH₂Cl₂ (21 mg, 0.10 equiv). Theresulting solution was stirred overnight at 80° C. in an oil bath. Thereaction mixture was cooled. The crude product was purified by Prep-HPLCproduct was obtained. This resulted in 23.9 mg (23%) of3-chloro-4-[6-chloro-2-(trifluoromethyl)-1H-indol-5-yl]benzonitrile as aoff-white solid. (ES, m/z): 353 [M−H]⁻; (300 MHz, DMSO-d6, ppm): δ 12.64(brs, 1H), 8.20 (s, 1H), 7.95-7.92 (m, 1H), 7.69 (s, 2H), 7.63-7.60 (d,J=8.1 Hz, 1H), 7.12 (s, 1H).

Compound number 356 (i.e.,6-chloro-5-(2-chloro-4-fluorophenyl)-2-(trifluoromethyl)-1H-indole) wasmade as follows.

Into a 40-mL round-bottom flask, was placed a solution of5-bromo-6-chloro-2-(trifluoromethyl)-1H-indole (100 mg, 0.34 mmol, 1.00equiv) in dioxane/H₂O (7 mL), (2-chloro-4-fluorophenyl)boronic acid(117.3 mg, 0.67 mmol, 2.00 equiv), K₃PO₄ (214 mg, 1.01 mmol, 3.00equiv), Pd(PPh₃)₄ (39 mg, 0.03 mmol, 0.10 equiv). The resulting solutionwas stirred overnight at 100° C. The crude product was purified byPrep-HPLC. This resulted in 41 mg (35%) of6-chloro-5-(2-chloro-4-fluorophenyl)-2-(trifluoromethyl)-1H-indole asbrown oil. (ES, m/z): [M−H]⁻ 346; ¹H NMR (300 MHz, DMSO): 12.57 (brs,1H), 7.65 (s, 2H), 7.60-7.56 (m, 1H), 7.46-7.41 (m, 1H), 7.35-7.28 (m,1H), 7.09 (s, 1H).

Compound number 357 (i.e.,6-Chloro-5-(2,4-dichlorophenyl)-2-(trifluoromethyl)-1H-indole) was madeas follows.

Into a 50-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed a solution of5-bromo-6-chloro-2-(trifluoromethyl)-1H-indole (100 mg, 0.34 mmol, 1.00equiv) in Dioxane/H₂O (6 mL/1 mL), (2,4-dichlorophenyl)boronic acid (128mg, 0.67 mmol, 2.00 equiv), K₃PO₄ (212 mg, 1.00 mmol, 3.00 equiv),Pd(PPh₃)₄ (39 mg, 0.03 mmol, 0.10 equiv). The resulting solution wasstirred overnight at 100° C. The resulting mixture was concentratedunder vacuum. The crude product was purified by Prep-HPLC. This resultedin 15.3 mg (13%) of6-chloro-5-(2,4-dichlorophenyl)-2-(trifluoromethyl)-1H-indole as brownoil. (ES, m/z): [M−H]⁻ 362; ¹H NMR (300 MHz, DMSO): 12.59 (br s, 1H),7.76 (d, J=2.1 Hz, 1H), 7.66 (s, 2H), 7.54-7.51 (m, 1H), 7.42 (d, J=8.4Hz, 1H), 7.10 (s, 1H).

Compound number 358 (i.e.,2-[6-Chloro-2-(trifluoromethyl)-1H-indol-5-yl]benzoic acid) was made asfollows.

Into a 50-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed a solution of5-bromo-6-chloro-2-(trifluoromethyl)-1H-indole (200 mg, 0.67 mmol, 1.00equiv) in Dioxane, H₂O (14 mL), 2-(dihydroxyboranyl)benzoic acid (223mg, 1.34 mmol, 2.00 equiv), K₃PO₄ (427 mg, 2.01 mmol, 3.00 equiv),Pd(PPh₃)₄ (78 mg, 0.07 mmol, 0.10 equiv). The resulting solution wasstirred overnight at 100° C. The crude product was purified byPrep-HPLC. This resulted in 10.2 mg (4%) of2-[6-chloro-2-(trifluoromethyl)-1H-indol-5-yl]benzoic acid as a whitesolid. (ES, m/z): [M−H]⁻ 338; ¹H NMR (300 MHz, DMSO): 12.43 (br s, 1H),7.96-7.91 (m, 1H), 7.65-7.60 (m, 1H), 7.56-7.49 (m, 3H), 7.31-7.29 (m,1H), 7.06 (s, 1H).

Compound number 359 (i.e.,2-[6-Chloro-2-(trifluoromethyl)-1H-indol-5-yl]benzonitrile) was made asfollows.

Into a 50-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed a solution of5-bromo-6-chloro-2-(trifluoromethyl)-1H-indole (100 mg, 0.34 mmol, 1.00equiv) in Dioxane, H₂O (7 mL), (2-cyanophenyl)boronic acid (99 mg, 0.67mmol, 2.00 equiv), K₃PO₄ (214 mg, 1.01 mmol, 3.00 equiv), Pd(PPh₃)₄ (39mg, 0.03 mmol, 0.10 equiv). The resulting solution was stirred overnightat 100° C. The resulting mixture was concentrated under vacuum. Thecrude product was purified by Prep-HPLC. This resulted in 17.4 mg (16%)of 2-[6-chloro-2-(trifluoromethyl)-1H-indol-5-yl]benzonitrile as a lightyellow solid. (ES, m/z): [M−H]⁻ 319; ¹H NMR (300 MHz, DMSO): 12.66 (brs, 1H), 7.96 (d, J=8.1 Hz, 1H), 7.79 (d, J=7.8 Hz, 2H), 7.71 (s, 1H),7.66 (d, J=7.5 Hz, 1H), 7.56 (d, J=8.1 Hz, 1H), 7.14 (s, 1H).

Compound number 360 (i.e.,([2-[6-Chloro-2-(trifluoromethyl)-1H-indol-5-yl]phenyl]methyl)dimethylamine)was made as follows.

Into a 50-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed a solution of5-bromo-6-chloro-2-(trifluoromethyl)-1H-indole (100 mg, 0.34 mmol, 1.00equiv) in Dioxane, H₂O (7 mL), 2-[(dimethylamino)methyl]phenylboronicacid (121 mg, 0.68 mmol, 2.00 equiv), K₃PO₄ (214 mg, 1.01 mmol, 3.00equiv), Pd(PPh₃)₄ (39 mg, 0.03 mmol, 0.10 equiv). The resulting solutionwas stirred overnight at 100° C. The resulting mixture was concentratedunder vacuum. The crude product was purified by Prep-HPLC. This resultedin 15.4 mg (13%) of([2-[6-chloro-2-(trifluoromethyl)-1H-indol-5-yl]phenyl]methyl)dimethylamineas a white solid. (ES, m/z): [M+H]⁺ 353; ¹H NMR (300 MHz, DMSO): 12.63(s, 1H), 9.66-9.59 (m, 1H), 7.80-7.69 (m, 3H), 7.60-7.51 (m, 2H),7.33-7.30 (m, 2H), 7.10 (s, 1H), 4.34-4.28 (m, 1H), 3.95-3.90 (m, 1H),2.73-2.64 (m, 3H), 2.51-2.46 (m, 3H).

Compound number 360-0A (i.e.,([2-[6-Chloro-2-(trifluoromethyl)-1H-indol-5-yl]phenyl]methyl)dimethylaminehydrochloride) was made as follows.

Into a 50-mL round-bottom flask, was placed a solution of5-bromo-6-chloro-2-(trifluoromethyl)-1H-indole (100 mg, 0.34 mmol, 1.00equiv) in Dioxane, H₂O (7 mL), 2-[(dimethylamino)methyl]phenylboronicacid (121 mg, 0.68 mmol, 2.00 equiv), K₃PO₄ (214 mg, 1.01 mmol, 3.00equiv), Pd(PPh₃)₄ (39 mg, 0.03 mmol, 0.10 equiv). The resulting solutionwas stirred overnight at 100° C. The resulting mixture was concentratedunder vacuum. The crude product was purified by Prep-HPLC. After that,conc HCl (aq, 1 mL) was added to the resulting solution and the solutionwas lyophilized. This resulted in 15.4 mg (12%) of([2-[6-chloro-2-(trifluoromethyl)-1H-indol-5-yl]phenyl]methyl)dimethylaminehydrochloride as a yellow solid. (ES, m/z): [M+H]⁺ 353; ¹H NMR (300 MHz,DMSO): 12.77 (br s, 1H), 10.45 (br s, 1H), 7.96 (d, J=7.5 Hz, 1H),7.81-7.70 (m, 2H), 7.58-7.52 (m, 2H), 7.39-7.24 (m, 1H). 7.19-7.10 (m,1H), 4.31-4.21 (m, 1H), 3.94-3.88 (m, 1H), 2.62 (d, J=4.5 Hz, 3H), 2.46(d, J=4.5 Hz, 3H).

Compound number 361 (i.e.,6-chloro-5-(3-chloropyridin-4-yl)-2-(trifluoromethyl)-1H-indole) wasmade as follows.

Into a 50-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed dioxane (10 mL), water (2 mL),4-bromo-3-chloropyridine (111 mg, 0.58 mmol, 1.00 equiv),6-chloro-5-(tetramethyl-1,3,2-dioxaborolan-2-yl)-2-(trifluoromethyl)-1H-indole(200 mg, 0.58 mmol, 1.00 equiv), sodium carbonate (122 mg, 1.14 mmol,2.00 equiv), tetrakis(triphenylphosphine) palladium (66 mg, 0.06 mmol,0.10 equiv). The resulting solution was stirred overnight at 80° C. inan oil bath. The reaction mixture was cooled. The crude product waspurified by Prep-HPLC. This resulted in 28.4 mg (15%) of6-chloro-5-(3-chloropyridin-4-yl)-2-(trifluoromethyl)-1H-indole as awhite solid. (ES, m/z): 329 [M−H]⁻; (300 MHz, DMSO-d6, ppm): δ 12.66(brs, 1H), 8.77 (s, 1H), 8.63 (d, J=4.8 Hz, 1H), 7.72 (d, J=6.3 Hz, 2H),7.49 (d, J=5.1 Hz, 1H), 7.13 (s, 1H).

Compound number 361-0A (i.e.,6-chloro-5-(3-chloropyridin-4-yl)-2-(trifluoromethyl)-1H-indolehydrochloride) was made as follows.

Into a 50-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed6-chloro-5-(3-chloropyridin-4-yl)-2-(trifluoromethyl)-1H-indole (25 mg,0.08 mmol, 1.00 equiv) and 10 ml water, 1 ml HCl. The combined aqueouslayer was dried by lyophilization. This resulted in 25.2 mg (91%) of6-chloro-5-(3-chloropyridin-4-yl)-2-(trifluoromethyl)-1H-indolehydrochloride as a white solid. (ES, m/z): 329 [M−HCl—H]⁻; (300 MHz,DMSO-d6, ppm): δ 8.80 (s, 1H), 8.65 (d, J=5.1 Hz, 1H), 7.72 (d, J=3.6Hz, 2H), 7.52 (d, J=4.8 Hz, 1H), 7.13 (s, 1H).

Compound number 362 (i.e.,6-chloro-5-(pyridin-4-yloxy)-2-(trifluoromethyl)-1H-indole;trifluoroacetic acid) was made as follows.

First, into a 100-mL 3-necked round-bottom flask was placedtetrahydrofuran (50 mL),6-chloro-5-(tetramethyl-1,3,2-dioxaborolan-2-yl)-2-(trifluoromethyl)-1H-indole(2.5 g, 7.23 mmol, 1.00 equiv), then added 15% NaOH adjust the pH=9, thereaction mixture stirred for 10 min at r.t. then added hydrogen peroxide(8.2 g, 241.07 mmol, 10.00 equiv). The resulting solution was stirredfor 2 h at room temperature. The resulting solution was diluted with 100mL of ethyl acetate. The resulting mixture was washed with 3×30 mL ofBrine. The mixture was dried over anhydrous sodium sulfate andconcentrated under vacuum. The residue was applied onto a silica gelcolumn with ethyl acetate/petroleum ether (1:30-1:20). This resulted in600 mg (35%) of 6-chloro-2-(trifluoromethyl)-1H-indol-5-ol as a yellowsolid.

Next, into a 50-mL round-bottom flask purged and maintained with aninert atmosphere of nitrogen, was placed dioxane (10 mL),6-chloro-2-(trifluoromethyl)-1H-indol-5-ol (230 mg, 0.98 mmol, 1.00equiv), (pyridin-4-yl)boronic acid (180 mg, 1.46 mmol, 1.50 equiv),(acetyloxy)cuprio acetate (53 mg, 0.29 mmol, 0.30 equiv), pyridine (231mg, 2.92 mmol, 3.00 equiv). The resulting solution was stirred overnightat 80° C. in an oil bath. The reaction mixture was cooled and the solidwas filtered. The crude product was purified by Prep-HPLC product wasobtained. This resulted in 15.8 mg (4%) of6-chloro-5-(pyridin-4-yloxy)-2-(trifluoromethyl)-1H-indole;trifluoroacetic acid as a yellow solid. (ES, m/z): 313 [MS-CF₃COOH+H]⁺;(300 MHz, DMSO-d6, ppm): δ 12.97 (s, 1H), 10.44 (s, 1H), 9.31 (d, J=5.4Hz, 2H), 8.87 (t, J=7.8 Hz, 1H), 8.41 (t, J=7.8 Hz, 2H), 8.90 (s, 1H),6.91 (s, 1H).

Compound number 362-0A (i.e.,6-chloro-5-(pyridin-4-yloxy)-2-(trifluoromethyl)-1H-indolehydrochloride) was made as follows.

Into a 50-mL round-bottom flask, was placed6-chloro-5-(pyridin-4-yloxy)-2-(trifluoromethyl)-1H-indole (15 mg, 0.05mmol, 1.00 equiv), 10 ml water and 1 ml HCl. The combined aqueous layerwas dried by lyophilization. This resulted in 16.7 mg of6-chloro-5-(pyridin-4-yloxy)-2-(trifluoromethyl)-1H-indole hydrochlorideas a grey solid. (ES, m/z): 313[Ms+H]⁺; (300 MHz, DMSO-d6, ppm): δ 13.08(s, 1H); 10.44 (s, 1H), 9.32 (d, J=2.4 Hz, 2H), 8.87 (t, J=8.7 Hz, 1H),8.41-8.36 (m, 2H), 7.91 (s, 1H), 6.90 (s, 1H).

Having thus described in detail various embodiments of the presentinvention, it is to be understood that the invention defined by theabove paragraphs is not to be limited to particular details set forth inthe above description as many apparent variations thereof are possiblewithout departing from the spirit or scope of the present invention.

What is claimed is:
 1. A compound of formula:

wherein: R₁ is (C₁-C₆)-alkyl substituted with one or more halogens; R₂is H, halogen, (C₁-C₆)-alkyl or (C₁-C₆)-haloalkyl; R₃ is biphenyl,naphthyl, aryl, aryl ether, or (C₃-C₆) cycloalkyl; wherein the lastthree substituents may be unsubstituted or substituted with one or moreof halogen, (C₁-C₃)-haloalkyl, (C₁-C₃)-haloalkoxy, aminoalkyl, cyano orcarboxyl; R₄ is halogen; R₅ is H, halogen, (C₁-C₆)-alkyl or(C₁-C₆)-haloalkyl; and R₆ is H, halogen, (C₁-C₆)-alkyl, (C₂-C₅)-alkenyl,or (C₂-C₅)-alkynyl, wherein each of (C₁-C₆)-alkyl, (C₂-C₅)-alkenyl, or(C₂-C₅)-alkynyl is independently unsubstituted or substituted with oneor more halogens.
 2. A compound according to claim 1 wherein R₁ is(C₁-C₄)-alkyl substituted with fluorine.
 3. A compound according toclaim 1 wherein R₂ is Cl.
 4. A compound according to claim 1 wherein R₃is a mono- or bi-halogen substituted phenyl ether.
 5. A compoundaccording to claim 1 wherein R₃ is a bi- or tri-halogen substitutedphenyl.
 6. A compound according to claim 4 wherein the halogen ischlorine, bromine or fluorine.
 7. A compound according to claim 5wherein the halogen is chlorine, bromine or fluorine.
 8. A compoundaccording to claim 1 wherein R₄ is chlorine.
 9. A compound according toclaim 1 wherein R₅ is hydrogen.
 10. A composition for treating helminthinfestation comprising an anthelmintically effective amount of thecompound of claim 1 and a pharmaceutically acceptable carrier.
 11. Acomposition for treating helminth infestation according to claim 10wherein the compound of formula (I) is combined with an additionalactive agent selected from the group consisting of acaricides,anthelmintics, anti-parasitics and insecticides.
 12. A composition fortreating helminth infestation according to claim 11 wherein the activeagent is a macrocyclic lactone.
 13. A composition for treating helminthinfestation according to claim 12 wherein the macrocyclic lactone isselected from the group consisting of abamectin, dimadectin, doramectin,emamectin, eprinomectin, ivermectin, latidectin, lepimectin, selamectinor ML-1,694,554.
 14. A composition for treating helminth infestationaccording to claim 10 wherein the compound of formula (I) is combinedwith verapamil.
 15. A method for treating helminth infestationcomprising administration of an anthelmintically effective amount of thecompound of claim 1 to an animal in need thereof.
 16. A method accordingto claim 15 in which the helminths are trematodes.
 17. A methodaccording to claim 16 in which the helminths are Fasciola hepatica. 18.A compound of formula:

wherein: R₁ is CF₃; R₂ is H; R₃ is phenyl or heteroaryl; wherein thephenyl or heteroaryl is unsubstituted or substituted with one or more ofhalogen, (C₁-C₃)-haloalkyl or cyano; R₄ is halogen; R₅ is H; and R₆ isH.
 19. A compound according to claim 18 wherein the heteroaryl is apyridine.
 20. A composition for treating helminth infestation comprisingan anthelmintically effective amount of the compound of claim 18 and apharmaceutically acceptable carrier.
 21. A composition for treatinghelminth infestation according to claim 20 wherein the compound offormula (I) is combined with an additional active agent selected fromthe group consisting of acaricides, anthelmintics, anti-parasitics andinsecticides.
 22. A composition for treating helminth infestationaccording to claim 21 wherein the active agent is a macrocyclic lactone.23. A composition for treating helminth infestation according to claim22 wherein the macrocyclic lactone is selected from the group consistingof abamectin, dimadectin, doramectin, emamectin, eprinomectin,ivermectin, latidectin, lepimectin, selamectin or ML-1,694,554.
 24. Acomposition for treating helminth infestation according to claim 20wherein the compound of formula (I) is combined with verapamil.
 25. Amethod for treating helminth infestation comprising administration of ananthelmintically effective amount of the compound of claim 18 to ananimal in need thereof.
 26. A method according to claim 25 in which thehelminths are trematodes.
 27. A method according to claim 26 in whichthe helminths are Fasciola hepatica.